Antitumoral analogs of lamellarins

ABSTRACT

New lamellarins are provided of the general formula III wherein X is selected from the group consisting of N, O and S; wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8  and R 9  are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SO 2 R′, NHR′, N(R′) 2 , N═R′, NHCOR′, N(COR′) 2 , NHS0 2 R′, N0 2 , PO(R′ 2 , P0 2 R′, C(=0)H, C(=0)R′, C0 2 H, C0 2 R′, OPO(R′) 2 , OP0 2 R′, OC(=0)H, OC(=0)R′, N═C(R′) 2 , substituted or unsubstituted C 1 -C 12  alkyl, substituted or unsubstituted C 1 -C 12  haloalkyl, substituted or unsubstituted C 2 -C 12  alkenyl, substituted or unsubstituted C 2 -C 12  alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and substituted or unsubstituted heteroaromatic; wherein each of the R′ groups is independently selected from the group consisting of H, OH, N0 2 , NH 2 , SH, CN, halogen, =0, C(=0)H, C(═O)CH 3 , C0 2 H, C(=0)R′, substituted or unsubstituted C 1 -C 18  alkyl, substituted or unsubstituted C 2 -C 18  alkenyl, substituted or unsubstituted C 2 -C 18  alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C 1 -C 18  alkoxyl, substituted or unsubstituted C 1 -C 18  aminoalkyl, substituted or unsubstituted C 1 -C 18  aminoacid, substituted or unsubstituted C 1 -C 18  thioalkyl, substituted or unsubstituted C 1 -C 18  alkylsulfinyl, substituted or unsubstituted C 1 -C 18  alkylsulfonyl; wherein the pairs of groups R 1  and R 2 , R 2  and R 3 , R 3  and R 4 , R 3  and R 9 , R 4  and R 9 , R 9  and R 5 , R 9  and R 6 , or R 6  and R 7 , R 7  and R 8  may be joined into a carbocyclic or heterocyclic ring system; and the dotted line represents an single or double bond; or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof.

FIELD OF THE INVENTION

The present invention relates to antitumoral compounds, and in particular to new antitumoral analogs of lamellarins, pharmaceutical compositions containing them and their use in the treatment of cancer.

BACKGROUND OF THE INVENTION

The lamellarins are polyaromatics alkaloids originally isolated from marine sources and comprising a fused polyaromatic framework. The family of lamellarins are constituted by two basic structures:

Both structures have a pyrrolic ring substituted with aryl units. The hexacyclic structure 1 is a 14-phenyl-6H-[1]benzopiran[4′,3′,4,5]pyrrolo[2,1-a]isoquinolin-6-one. Depending of the substituents and the presence of a double bond between C₈-C₉ the members of this family are designed with different letters.

R₁ R₂ R₃ R₄ R₅ R₆ R₇ R₈ A OCH₃ CH₃ CH₃ H CH₃ CH₃ H OH C OCH₃ CH₃ CH₃ H CH₃ CH₃ H H E OH CH₃ CH₃ CH₃ H CH₃ H H F OH CH₃ CH₃ CH₃ CH₃ CH₃ H H G H H CH₃ CH₃ H H CH₃ H I OCH₃ CH₃ CH₃ CH₃ CH₃ CH₃ H H J H H CH₃ CH₃ CH₃ CH₃ H H K OH CH₃ CH₃ H CH₃ CH₃ H H L H H CH₃ CH₃ H CH₃ H H S H H CH₃ H H H H H T OCH₃ CH₃ CH₃ CH₃ H CH₃ H H U H CH₃ CH₃ CH₃ H CH₃ H H V OCH₃ CH₃ CH₃ CH₃ H CH₃ H OH Y H CH₃ H CH₃ H CH₃ SO₃Na H Z H H CH₃ H H H CH₃ H β H H H CH₃ H H H H

R₁ R₂ R₃ R₄ R₅ R₆ R₇ B OCH₃ CH₃ CH₃ H CH₃ CH₃ H D H H CH₃ H CH₃ CH₃ H H H H H H H H H M OH CH₃ CH₃ H CH₃ CH₃ H N H H CH₃ CH₃ H CH₃ H W OCH₃ CH₃ CH₃ CH₃ H CH₃ H X OH CH₃ CH₃ CH₃ H CH₃ H α H CH₃ CH₃ CH₃ H CH₃ SO₃Na

R. J. Anderson et al, J. Am. Chem. Soc. 1985, 107, 5492, describes the isolation and characterization of four polyaromatic metabolites, the lamellarins A-D, obtained from a marine prosobranch mollusc Lamellana sp. The structure of lamellarin A was determined by an X-Ray crystallographic study and the structures of lamellarins B-D were assigned by interpretation of spectral data.

N. Lindquist et al, J. Org. Chem. 1988, 53, 4570, describes the isolation and characterization of four new lamellarins: E-H from the marine ascidian Didemnum chartaceum obtained from the Indian Ocean. The structure of lamellarin E was determined by an X-Ray crystallographic study.

A. R. Carroll et al, Aust. J. Chem. 1993, 46, 489, isolated six new lamellarins: I, J, K, L, M and the triacetate of the lamellarin N, and four known of this type: A, B, C, and the triacetate of lamellarin D, isolated from a marine ascidian Didemnum sp.

S. Urban et al, Aust. J. Chem. 1994, 47, 1919 and Aust. J. Chem. 1995, 48, 1491, described the isolation and characterization of four new lamellarins, O, P, Q, R, with the substructure type 2 from the marine sponge Dendnilla cactos. Later S. Urban et al, Aust. J. Chem. 1996, 49, 711, described the structure of lamellarin S from the ascidian Didemnum sp.

M. V. R. Reddy et al, Tetrahedron 1997, 53, 3457, isolated five new lamellarins: T, U, V, W, and X, and the first example of sulfated lamellarin, Y, isolated from the marine ascidian Didemnum sp obtained from the Arabian sea.

R. A. Davis et al, J. Nat. Prod. 1999, 62, 419, described one new lamellarin, Z, and various examples of sulphated lamellarins isolated from the marine ascidian Didemnum chartaceum.

M. V. R. Reddy et al, J. Med. Chem. 1999, 42, 1901, isolated a new lamellarin, α, isolated from the marine ascidian Didemnum sp.

Finally, J. Ham et al, Bull. Korean Chem. Soc. 2002, 23, 163, described the isolation and characterization of the lamellarin β obtained from a marine ascidian Didemnum sp.

Lamellarins C and D have been shown to cause inhibition of cell division in a fertilised sea urchin assay, whereas lamellarins I, K, and L all exhibit comparable cytoxicity against P388 and A549 cell lines in culture. Recently, lamellarin N has been shown to exhibit activity in lung cancer cell lines by acting as a Type IV microtubule poison.

Furthermore, J. L. Fernández-Puentes et al, PCT Int. Appl WO 97/01336, describe that these compounds have also cytotoxic activity on multidrug resistant cells as well as efficacy as non-toxic modulators of the multidrug resistant phenotype and, therefore, afford an attractive potential source of chemotherapeutic agents.

The limited availability of natural material has resulted in the search for alternative synthetic methods being sought for the natural compounds and related analogs. M. G. Banwell et al, Int. Patent Appl. WO 98/50365 and Int. Patent Appl. WO 99/67250 described the synthesis of lamellarin K via 1,3-dipolar cycloaddition between an alkyne and an N-ylide of isoquinolin.

Lamellarin G trimethyl ether was also synthetised by S. Ruchirawat et al, Tetrahedron Lett. 2001, 42, 1250. The synthesis involved the formation of the core pyrrolo[2,1-a]isoquinoline, followed by the formation of the lactone ring.

Lamellarins I and K (1) were obtained by L. Castedo et al, Synlett 2001, 7, 1164, by a new approach based on the 1,3-dipolar cycloaddition of a nitrone to an alkyne. The key cycloaddition yield an isoxazoline which rearranged to afford the central pyrrole ring.

F. Albericio et al, Org. Lett 2003, 5, 2959, has described a total solid-phase synthesis of Lamellarins U and L.

Ishibashi F. et al., J. Nat. Prod., 2002, 65, 500-504 describe the synthesis and structure activity relationship for some lamellarin derivatives.

The discovery of the main target for an anticancer agent is an essential element to better understand its mechanism of action and to guide the development of clinically useful analogs. To illustrate this, one can refer to camptothecin (CPT) discovered in the early 1960s but successfully developed only a quarter of a century later when its main, and perhaps unique, molecular target was identified: topoisomerase I. The observation in 1985 that CPT stabilizes DNA-topoisomerase I complexes provided the starting point for the rational development of safe CPT analogs which culminated in the mid 1990s with the approval of topotecan and irinotecan for the treatment of ovarian and colon cancers.

The search for non-CPT topoisomerase I poisons has been very active for the past ten years but only a limited number of potent topoisomerase I poisons has been discovered.

SUMMARY OF THE INVENTION

We have found that the lamellarins represent a new and promising series of topoisomerase I inhibitors. The correlation between the capacity of the drugs to stimulate topoisomerase I-mediated DNA cleavage and their cytotoxic potential makes them useful as antitumor agents.

The present invention is directed to compounds of the general formula III:

wherein X is selected from the group consisting of N, O and S; wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO₂R′, NHR′, N(R′)₂, N═R′, NHCOR′, N(COR′)₂, NHSO₂R′, NO₂, PO(R′)₂, PO₂R′, C(═O)H, C(═O)R′, CO₂H, CO₂R′, OPO(R′)₂, OPO₂R′, OC(═O)H, OC(═O)R′, N═C(R′)₂, substituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₁-C₁₂ haloalkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, substituted or unsubstituted C₂-C₁₂ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and substituted or unsubstituted heteroaromatic; wherein each of the R′ groups is independently selected from the group consisting of H, OH, NO₂, NH₂, SH, CN, halogen, ═O, C(═O)H, C(═O)CH₃, CO₂H, C(═O)R′, substituted or unsubstituted C₁-C₁₈ alkyl, substituted or unsubstituted C₂-C₁₈ alkenyl, substituted or unsubstituted C₂-C₁₈ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C₁-C₁₈ alkoxyl, substituted or unsubstituted C₁-C₁₈ aminoalkyl, substituted or unsubstituted C₁-C₁₈ aminoacid, substituted or unsubstituted C₁-C₁₈ thioalkyl, substituted or unsubstituted C₁-C₁₈ alkylsulfinyl, substituted or unsubstituted C₁-C₁₈ alkylsulfonyl; wherein the pairs of groups R₁ and R₂, R₂ and R₃, R₃ and R₄, R₃ and R₉, R₄ and R₉, R₉ and R₅, R₉ and R₆, or R₆ and R₇, R₇ and R₅ may be joined into a carbocyclic or heterocyclic ring system; and the dotted line represents an single or double bond; or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof.

We exclude compounds that are known lamellarins, especially known lamellarins described in the literature acknowledged in the present introduction, and more especially lamellarins A-N and S-Z or lamellarins α or β, as well as lamellarin D, K, L, M or N triacetate, lamellarin G trimethyl ether and compounds in WO 9850365. In this respect, we explicitly incorporate by specific reference each of the prior art documents mentioned in the present introduction, particularly for any disclsoure of a known compound which needs to be excluded from the repsent claims.

PREFERRED EMBODIMENTS

Preferred compounds of this invention are those of formula IV:

wherein R₁-R₈ are as defined above and R′₂-R′₆ and the dotted line have the same definitions as for R₁-R₈. In this respect, appropriate pairs of R′₂-R′₆ may be joined into a carbocyclic or heterocyclic ring system.

In formula III or IV X is preferably O or NH, most preferably O.

Preferred compounds are those that have a double bond between C-8 and C-9 (dotted line), they have shown higher antitumoral activity.

In a preferred aspect of the invention each of R₁-R₈ is independently selected from H, OR′, OC(═O)R′.

R₃ is preferably selected from the group consisting of H, OH, alkoxy, most preferably methoxy.

R₄, R₅, R₆ and R₈ are preferably each independently selected from the group consisting of H or alkoxy, most preferably they are H. Suitably at least 2, 3 or preferably all 4 of R₄, R₅, R₆ and R₈ are the same, and R₃ is preferably that group.

R₁, R₂ and R₇ are preferably each independently selected from the group consisting of H, OH, alkoxy, OC(═O)R′, SO₂R′, PO(R′)₂, Alkyl, NO₂, NH₂.

In a most preferred embodiment R₁, R₂ and R₇ are OC(═O)R′ wherein R′ is a substituted or unsubstituted aminoacid or aminoacids chain, preferably with a cationic group. Suitably at least 2, or preferably all 3 of R₁, R₂ and R₇ are the same.

In formula IV R′₂, R′₃ and R′₆ are preferably each independently selected from the group consisting of H or alkoxy, most preferably H; and R′₅ is preferably selected from the group consisting of H or alkoxy, most preferably methoxy.

R′₄ is preferably selected from the group consisting of H, OH, alkoxy, OC(═O)R′, SO₂R′, PO(R′)₂, Alkyl, NO₂, NH₂. Most preferably R′₄ is C(═O)R′ wherein R′ is a substituted or unsubstituted aminoacid or aminoacids chain, preferably with a cationic group.

Often R′₄, R₇ and either R₁ or R₂ is the same.

Any of the groups with a protectable hydroxy or amino subsituent may be in protected form, using available protecting groups.

Suitable protecting groups for phenols and hydroxy groups include ethers and esters, such as alkyl, alkoxyalkyl, aryloxyalkyl, alkoxyalkoxyalkyl, alkylsilylalkoxyalkyl, alkylthioalkyl, arylthioalkyl, azidoalkyl, cyanoalkyl, chloroalkyl, heterocyclic, arylacyl, haloarylacyl, cycloalkylalkyl, alkenyl, cycloalkyl, alyklarylalkyl, alkoxyarylalkyl, nitroarylalkyl, haloarylalkyl, alkylaminocarbonylarylalkyl, alkylsulfinylarylalkyl, alkylsilyl and other ethers, and arylacyl, aryl alkyl carbonate, aliphatic carbonate, alkylsulfinylarlyalkyl carbonate, alkyl carbonate, aryl haloalkyl carbonate, aryl alkenyl carbonate, aryl carbamate, alkyl phosphinyl, alkylphosphinothioyl, aryl phosphinothioyl, aryl alkyl sulphonate and other esters. Such groups may optionally be substituted with the previously mentioned groups in R¹.

Suitable protecting groups for amines include carbamates, amides, and other protecting groups, such as alkyl, arylalkyl, sulpho- or halo-arylalkyl, haloalkyl, alkylsilylalkyl, arylalkyl, cycloalkylalkyl, alkylarylalkyl, heterocyclylalkyl, nitroarylalkyl, acylaminoalkyl, nitroaryldithioarylalkyl, dicycloalkylcarboxamidoalkyl, cycloalkyl, alkenyl, arylalkenyl, nitroarylalkenyl, heterocyclylalkenyl, heterocyclyl, hydroxyheterocyclyl, alkyldithio, alkoxy- or halo- or alkylsulphinyl arylalkyl, hetercyclylacyl, and other carbamates, and alkanoyl, haloalkanoyl, arylalkanoyl, alkenoyl, heterocyclylacyl, aroyl, arylaroyl, haloaroyl, nitroaroyl, and other amides, as well as alkyl, alkenyl, alkylsilylalkoxyalkyl, alkoxyalkyl, cyanoalkyl, heterocyclyl, alkoxyarylalkyl, cycloalkyl, nitroaryl, arylalkyl, alkoxy- or hydroxy-arylalkyl, and many other groups. Such groups may optionally be substituted with the previously mentioned groups in R¹.

Examples of such protecting groups are given in the following tables. protection for —OH group abbreviation ethers methyl methoxymethyl MOM benzyloxymethyl BOM methoxyethoxymethyl MEM 2-(trimethylsilyl) ethoxymethyl SEM methylthiomethyl MTM phenylthiomethyl PTM azidomethyl cyanomethyl 2,2-dichloro-1,1-difluoroethyl 2-chloroethyl 2-bromoethyl tetrahydropyranyl THP 1-ethoxyethyl EE phenacyl 4-bromophenacyl cyclopropylmethyl allyl propargyl isopropyl cyclohexyl t-butyl benzyl 2,6-dimethylbenzyl 4-methoxybenzyl MPM or PMB o-nitrobenzyl 2,6-dichlorobenzyl 3,4-dichlorobenzyl 4-(dimethylamino)carbonylbenzyl 4-methylsuflinylbenzyl Msib 9-anthrylmethyl 4-picolyl heptafluoro-p-tolyl tetrafluoro-4-pyridyl trimethylsilyl TMS t-butyldimethylsilyl TBDMS t-butyldiphenylsilyl TBDPS triisopropylsilyl TIPS esters aryl formate aryl acetate aryl levulinate aryl pivaloate ArOPv aryl benzoate aryl 9-fluorocarboxylate aryl methyl carbonate 1-adamantyl carbonate t-butyl carbonate BOC-OAr 4-methylsulfinylbenzyl carbonate Msz-Oar 2,4-dimethylpent-3-yl carbonate Doc-Oar aryl 2,2,2-trichloroethyl carbonate aryl vinyl carbonate aryl benzyl carbonate aryl carbamate dimethylphosphinyl Dmp-OAr dimethylphosphinothioyl Mpt-OAr diphenylphosphinothioyl Dpt-Oar aryl methanesulfonate aryl toluenesulfonate aryl 2-formylbenzenesulfonate

protection for the —NH₂ group abbreviation carbamates methyl ethyl 9-fluorenylmethyl Fmoc 9-(2-sulfo)fluroenylmethyl 9-(2,7-dibromo)fluorenylmethyl 17-tetrabenzo[a,c,g,i]fluorenylmethyl Tbfmoc 2-chloro-3-indenylmethyl Climoc benz[f]inden-3-ylmethyl Bimoc 2,7-di-t-butyl[9-(10,10-dioxo-10,10,10,10- DBD-Tmoc tetrahydrothioxanthyl)]methyl 2,2,2-trichloroethyl Troc 2-trimethylsilylethyl Teoc 2-phenylethyl hZ 1-(1-adamantyl)-1-methylethyl Adpoc 2-chlooethyl 1,1-dimethyl-2-chloroethyl 1,1-dimethyl-2-bromoethyl 1,1-dimethyl-2,2-dibromoethyl DB-t-BOC 1,1-dimethyl-2,2,2-trichloroethyl TCBOC 1-methyl-1-(4-biphenyl)ethyl Bpoc 1-(3,5-di-t-butylphenyl)-1-1-methylethyl t-Burmeoc 2-(2′-and 4′-pyridyl)ethyl Pyoc 2,2-bis(4′-nitrophenyl)ethyl Bnpeoc n-(2-pivaloylamino)-1,1-dimethylethyl 2-[(2-nitrophenyl) dithio]-1-phenylethyl NpSSPeoc 2-(n,n-dicyclohexylcarboxamido)ethyl t-butyl BOC 1-adamantyl 1-Adoc 2-adamantyl 2-Adoc vinyl Voc allyl Aloc or Alloc 1-isopropylallyl Ipaoc cinnamyl Coc 4-nitrocinnamyl Noc 3-(3′-pyridyl)prop-2-enyl Paloc 8-quinolyl n-hydroxypiperidinyl alkyldithiobenzyl Cbz or Z p-methoxybenzyl Moz p-nitrobenzyl PNZ p-bromobenzyl p-chlorobenzyl 2,4-dichlorobenzyl 4-methylsulfinylbenzyl Msz 9-anthrylmethyl diphenylmethyl phenothiazinyl-(10)-carbonyl n′-p-toluenesulfonylaminocarbonyl n′-phenylaminothiocarbonyl amides formamide acetamide chloroacetamide trifluoroacetamide TFA phenylacetamide 3-phenylpropanamide pent-4-enamide picolinamide 3-pyridylcarboxamide benzamide p-phenylbenzamide n-phthalimide n-tetrachlorophthalimide TCP 4-nitro-n-phthalimide n-dithiasuccinimide Dts n-2,3-diphenylmaleimide n-2,5-dimethylpyrrole n-2,5-bis(triisopropylsiloxyl)pyrrole BIPSOP n-1,1,4,4- STABASE tetramethyldisiliazacyclopentante adduct 1,1,3,3-tetramethyl-1,3-disilaisoindoline BSB special —NH protective groups n-methylamine n-t-butylamine n-allylamine n-[2-trimethylsilyl)ethoxy]methylamine SEM n-3-acetoxypropylamine n-cyanomethylamine n-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3- yl)amine n-2,4-dimethoxybenzylamine Dmb 2-azanorbornenes n-2,4-dinitrophenylamine n-benzylamine Bn n-4-methoxybenzylamine MPM n-2,4-dimethoxybenzylamine DMPM n-2-hydroxybenzylamine Hbn n-(diphenylmethyl)amino DPM n-bis(4-methoxyphenyl)methylamine n-5-dibenzosuberylamine DBS n-triphenylmethylamine Tr n-[(4- MMTr methoxyphenyl)diphenylmethyl]amino n-9-phenylflurenylamine Pf n-ferrocenylmethylamine Fcm n-2-picolylamine n′-oxide n-1,1-dimethylthiomethyleneamine n-benzylideneamine n-p-methoxybenzylideneamine n-diphenylmethyleneamine n-(5,5-dimethyl-3-oxo-1- cyclohexenyl)amine n-nitroamine n-nitrosoamine diphenylphosphinamide Dpp dimethylthiophosphinamide Mpt diphenylthiophosphinamide Ppt dibenzyl phosphoramidate 2-nitrobenzenesulfenamide Nps n-1-(2,2,2-trifluoro-1,1- TDE diphenyl)ethylsufenamide 3-nitro-2-pyridinesulfenamide Npys p-toluenesulfonamide Ts benzenesulfonamide

It is preferred that at least one of R₁-R₈ and R′₂-R′₆ is not H, OH, OCH₃, SO₃Na, most preferably at least two are not H, OH, OCH₃, SO₃Na. It is also preferred that at least one of these substituents has at least 2, more preferably at 3, yet more preferably at least 4 carbon atoms. In particular, we prefer that R′₄ and R₇, and possibly also R₁, have these minimal numbers of carbon atoms.

Antitumoral activities of these compounds include leukaemias, lung cancer, colon cancer, kidney cancer, prostate cancer, pancreatic cancer, cervix cancer, ovarian cancer, breast cancer, sarcomas and melanomas.

In another aspect the present invention is directed to pharmaceutical compositions useful as antitumor agents that contain as active ingredient a compound or compounds of the invention or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof and a pharmaceutically acceptable carrier.

The present invention is also directed to the use compounds of the general formula III above or pharmaceutically acceptable salts, derivatives, prodrugs or stereoisomers thereof in the treatment of cancer, or in the preparation of a medicament for the treatment of cancer.

In a further aspect the present invention is also directed to the use of compounds of the general formula III above or pharmaceutically acceptable salts, derivatives, prodrugs or stereoisomers thereof as topoisomerase I inhibitors.

DETAILED DESCRIPTION OF THE INVENTION

Fifteen years of efforts in targeting topoisomerase I for the discovery of anticancer agents have lead to the identification of several families of compounds capable of stabilizing DNA-topoisomerase I covalent complexes. The lead series is with no doubt the camptothecin family with two drugs, topotecan and irinotecan, approved for cancer treatment and several second (e.g. lurtotecan, exatecan) and third (e.g. diflomotecan) generations of camptothecin analogs currently in clinical trials. However, apart from the camptothecins, only a few topoisomerase I poisons have reached phase I clinical trials. Promising results have been reported with glycosyl indolocarbazoles but so far there is still no non-CPT topoisomerase I poisons in advanced clinical trials. The need for new series of topoisomerase I poisons remains pressing.

We have now surprisingly found that the natural lamellarins and their analogs are potent Topoisomerase I inhibitors, and that they exhibit sequence specificity profiles distinct from Camptothecin which is a well know Topoisomerase I inhibitor and chemotherapeutic agent, suggesting that they recognize differently with the topoisomerase I-DNA interface.

Therefore the invention is directed at compounds of formula III as defined above, their use as antitumoral agents and pharmaceutical compositions containing them.

The following gives guidance for the substituents in formulae III and IV:

Preferred R′ groups are present in groups of formula R′, COR′ or OCOR′ and include alkyl or alkenyl, that may be substituted at one or more available positions by one or more suitable groups, e.g., halogen such as fluoro, chloro, bromo and iodo, especially ω-chloro or perfluoro; aminoalkyl groups such as groups having one or more N atoms and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms, and especially including substituted or unsubstituted aminoacids or aminoacid chains, notably glycine, alanine, arginine, asparagine, asparaginic acid, cystein, glutamine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tiyptophan, tyrosine or valine, especially protected forms of such amino acids; carbocylic aryl having 6 or more carbons, particularly phenyl; and aralkyl such as benzyl; heterocyclic groups including heteroalicyclic and heteroaromatic groups, especially with 5 to 10 ring atoms of which 1 to 4 are heteroatoms, more preferably heterocyclic groups with 5 or 6 ring atoms and 1 or 2 heteratoms or with 10 ring atoms and 1 to 3 heteroatoms, the heterocyclic groups optionally being substituted with one or more of the subsitituents permitted for R′ and especially amino such as dimethylamino or with keto.

Suitable halogen substituents in the compounds of the present invention include F, Cl, Br and I.

Alkyl groups preferably have from 1 to 24 carbon atoms. One more preferred class of alkyl groups has 1 to about 12 carbon atoms, yet more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Another more preferred class of alkyl groups has 12 to about 24 carbon atoms, yet more preferably 12 to about 18 carbon atoms, and most preferably 13, 15 or 17 carbon atoms. Methyl, ethyl and propyl including isopropyl are particularly preferred alkyl groups in the compounds of the present invention. As used herein, the term alkyl, unless otherwise modified, refers to both cyclic and noncyclic groups, although cyclic groups will comprise at least three carbon ring members.

Preferred alkenyl and alkynyl groups in the compounds of the present invention have one or more unsaturated linkages and from 2 to about 12 carbon atoms, more preferably 2 to about 8 carbon atoms, still more preferably 2 to about 6 carbon atoms, even more preferably 2, 3 or 4 carbon atoms. The terms alkenyl and alkynyl as used herein refer to both cyclic and noncyclic groups, although straight or branched noncyclic groups are generally more preferred.

Alkylidene groups may be branched or unbranched and preferably have from 1 to 12 cabon atoms. One more preferred class of alkylidene groups has from 1 to about 8 carbon atoms, yet more preferably from 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Methylidene, ethylidene and propylidene including isopropylidene are particularly preferred alkylidene groups in the compounds of the present invention.

Preferred alkylsulfinyl groups in the compounds of the present invention include those groups having one or more sulfoxide (SO) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylsulfinyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.

Preferred alkylsulfonyl groups in the compounds of the present invention include those groups having one or more sulfonyl (SO₂) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylsulfonyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.

Preferred aminoalkyl groups include those groups having one or more primary, secondary and/or tertiary amine groups, and from 1 to about 12 carbon atoms, more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, even more preferably 1, 2, 3 or 4 carbon atoms. Secondary and tertiary amine groups are generally more preferred than primary amine moieties.

Suitable heterocyclic groups include heteroaromatic and heteroalicyclic groups. Suitable heteroaromatic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., coumarinyl including 8-coumarinyl, quinolinyl including 8-quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazol. Suitable heteroalicyclic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino and pyrrolindinyl groups.

Suitable carbocyclic aryl groups in the compounds of the present invention include single and multiple ring compounds, including multiple ring compounds that contain separate and/or fused aryl groups. Typical carbocyclic aryl groups contain 1 to 3 separate or fused rings and from 6 to about 18 carbon ring atoms. Specifically preferred carbocyclic aryl groups include phenyl including substituted phenyl such as 2-substituted phenyl, 3-substituted phenyl, 2.3-substituted phenyl, 2.5-substituted phenyl, 2.3.5-substituted and 2.4.5-substituted phenyl, including where one or more of the phenyl substituents is an electron-withdrawing group such as halogen, cyano, nitro, alkanoyl, sulfinyl, sulfonyl and the like; naphthyl including 1-naphthyl and 2-naphthyl; biphenyl; phenanthryl; and anthracyl.

References herein to substituted R′ groups in the compounds of the present invention refer to the specified moiety, typically alkyl or alkenyl, that may be substituted at one or more available positions by one or more suitable groups, e.g., halogen such as fluoro, chloro, bromo and iodo; cyano; hydroxyl; nitro; azido; alkanoyl such as a C₁₋₆ alkanoyl group such as acyl and the like; carboxamido; alkyl groups including those groups having 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms and more preferably 1-3 carbon atoms; alkenyl and alkynyl groups including groups having one or more unsaturated linkages and from 2 to about 12 carbon or from 2 to about 6 carbon atoms; alkoxy groups having those having one or more oxygen linkages and from 1 to about 12 carbon atoms or 1 to about 6 carbon atoms; aryloxy such as phenoxy; alkylthio groups including those moieties having one or more thioether linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; alkylsulfinyl groups including those moieties having one or more sulfinyl linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; alkylsulfonyl groups including those moieties having one or more sulfonyl linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; aminoalkyl groups such as groups having one or more N atoms and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; carbocylic aryl having 6 or more carbons, particularly phenyl (e.g., R being a substituted or unsubstituted biphenyl moiety); and aralkyl such as benzyl; heterocyclic groups including heteroalicyclic and heteroaromatic groups, especially with 5 to 10 ring atoms of which 1 to 4 are heteroatoms, more preferably heterocyclic groups with 5 or 6 ring atoms and 1 or 2 heteratoms or with 10 ring atoms and 1 to 3 heteroatoms.

As further guidance, we prefer as substituents for R₁-R₉ and R′₂-R′₆:

Amino Acids and Peptides

(L)-Val-OH; (L)-N-Boc-Val-OH

(D)-Val-OH; (D)-N-Boc-Val-OH

(L)-Ala-OH; (L)-N-Boc-Ala-OH; (L)-N-Alloc-Ala-OH; (L)-N-Fmoc-Ala-OH

(L)-Phe-OH; (L)-N-Boc-Phe-OH

(L)-N-Boc-Lys(Cbz)-OH

(L)-Leu-OH; (L)-N-Boc-Leu-OH

(L)-Pro-OH; (L)-N-Boc-Pro-OH

(L)-Trp-OH; (L)-N-Boc-Trp-OH

(L)-Ile-OH; (L)-N-Boc-Ile-OH

(L)-Ser(Bn)-OH; (L)-N-Boc-Ser(Bn)-OH

(L)-Cys(Fm)-OH; (L)-N-Boc-Cys(Fm)-OH

(L)-N-Boc-β-Leu-OH

(L)-N-Boc-Lys(Boc) Gly-OH

(L)-AlaAla-OH; (L)-N-Boc-AlaAla-OH

Esters

Hydrocinnamoyl

Cyclohexylpropyl

Methanosulfonyl (Ms)

Trifluoromethanosulfonyl (Tf)

Octanoyl

Biotin

Acetyl

Coumarin 3-carboxyl

2 [(4-fluorophenyl)thio]acetyl

4-fluorenecarboxyl

9H-fluorene-4-carboxyl

2,3,4,5-Tetrafluorobenzoyl

4-Pentynoyl

4-Methyl cinnamoyl

3,5-Dibromobenzoyl

5(2-Phenyleth-1-ynyl)nicotinyl

6-(Boc-amino)caproyl

6-Aminocaproyl

3-(Boc-amino)propyl

3-Aminopropyl

Ethers

Methyl

Isopropyl

Benzyl

4-Methoxybenzyl

Methoxymethyl

Methilenedioxy

Tert-butyldiphenylsilyl

Nitrogen Compounds

Nitro

Amino

Methylamino

Dimethylamino

Benzophenone imine

Phosphates

Diethyl phosphate

Halogens

Cl, Br, I

Cyanides

CN

The term “pharmaceutically acceptable salts, derivatives, prodrugs” refers to any pharmaceutically acceptable salt, ester, solvate, hydrate or any other compound which, upon administration to the recipient is capable of providing (directly or indirectly) a compound as described herein. However, it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts. The preparation of salts, prodrugs and derivatives can be carried out by methods known in the art.

For instance, pharmaceutically acceptable salts of compounds provided herein are synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two. Generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Examples of the acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulphate, nitrate, phosphate, and organic acid addition salts such as, for example, acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulphonate and p-toluenesulphonate. Examples of the alkali addition salts include inorganic salts such as, for example, sodium, potassium, calcium and ammonium salts, and organic alkali salts such as, for example, ethylenediamine, ethanolamine, N,N-dialkylenethanolamine, triethanolamine and basic aminoacids salts.

The compounds of the invention may be in crystalline form either as free compounds or as solvates (e.g. hydrates) and it is intended that both forms are within the scope of the present invention. Methods of solvation are generally known within the art.

Any compound that is a prodrug of a compound of formula III is within the scope and spirit of the invention. The term “prodrug” is used in its broadest sense and encompasses those derivatives that are converted in vivo to the compounds of the invention. Such derivatives would readily occur to those skilled in the art, and include, for example, compounds where a free hydroxy group is converted into an ester derivative.

The compounds of the present invention represented by the above described formula III may include enantiomers depending on their asymmetry or diastereoisomers. The single isomers and mixtures of the isomers fall within the scope of the present invention.

The compound of the present invention can be prepared synthetically from the intermediate compound Va described in the PCT Int. Appl WO 98 50365. Numerous active antitumoral compounds have been prepared from this compound and it is believed that many more compounds can be formed in accordance with the teachings of the present disclosure.

The compounds of formula III can be prepared from simple starting materials based on the following retrosynthesis.

Dependent on the choice of the R substituents of the starting materials or the chemical transformations into different definitions of this R, the methodology can provide access to a wide range of Lamellarins analogs as exemplified herein.

The preparation of compounds of general formula III is illustrated below for R₉ as H, Halogen, substituted or unsubstituted aryl or substituted or unsubstituted heteroaromatic.

The preparation of compounds of general formula IV is illustrated below:

Further details are given in the the experimental procedures and the physicochemical characteristics of the compounds in the examples.

Another especially preferred embodiment of the present invention is pharmaceutical compositions useful as antitumor agents which contain as active ingredient a compound or compounds of the invention, as well as the processes for their preparation.

An important feature of the above described compounds of formula III is their bioactivity and in particular their cytotoxic activity. With this invention we provide novel pharmaceutical compositions of compounds of general formula III that possess cytotoxic activity, and their use as antitumor agents. Thus the present invention further provides pharmaceutical compositions comprising a compound of this invention, a pharmaceutically acceptable salts, derivatives, prodrugs or stereoisomers thereof with a pharmaceutically acceptable carrier.

Examples of pharmaceutical compositions include any solid (tablets, pills, capsules, granules etc.) or liquid (solutions, suspensions or emulsions) with suitable composition for oral, topical or parenteral administration.

Administration of the compounds or compositions of the present invention may be any suitable method, such as intravenous infusion, oral preparation, intraperitoneal and intravenous preparation. We prefer that infusion times of up to 24 hours are used, more preferably 2-12 hours, with 2-6 hours most preferred. Short infusion times which allow treatment to be carried out without an overnight stay in hospital are especially desirable. However, infusion may be 12 to 24 hours or even longer if required. Infusion may be carried out at suitable intervals of say 1 to 4 weeks. Pharmaceutical compositions containing compounds of the invention may be delivered by liposome or nanosphere encapsulation, in sustained release formulations or by other standard delivery means.

The correct dosage of the compounds will vary according to the particular formulation, the mode of application, and the particular situs, host and tumour being treated. Other factors like age, body weight, sex, diet, time of administration, rate of excretion, condition of the host, drug combinations, reaction sensitivities and severity of the disease shall be taken into account. Administration can be carried out continuously or periodically within the maximum tolerated dose.

The compounds and compositions of this invention may be used with other drugs to provide a combination therapy. The other drugs may form part of the same composition, or be provided as a separate composition for administration at the same time or a different time.

Antitumoral activities of these compounds include among others leukaemias, lung cancer, colon cancer, kidney cancer, prostate cancer, ovarian cancer, breast cancer, pancreas cancer, cervix cancer, sarcomas and melanomas.

The present invention will be further explained with the following examples. These examples are illustrative of the present invention and should not not be interpreted as limitative.

EXAMPLES Example 1 Synthesis of Compounds 1-240

General Procedure A

A 0.15M suspension of the corresponding Isopropoxylated-Lamellarin (1 eq.) and AlCl₃ (1.3 eq. per isopropoxy group) in anhydrous dichloromethane was stirred at room temperature until the reaction was completed (2 to 6 h) under Argon atmosphere. Methanol was added, the solvent was evaporated under reduced pressure and the residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure B

A solution of anhydrous dichloromethane/TFA (3:1) was added to the corresponding Boc-aminoacid-Lamellarin (0.01M) at 0° C. under Argon atmosphere. The reaction mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure and the mixture was treated with dichloromethane in order to remove the remaining TFA. After final evaporation to dryness, the corresponding Lamellarin was collected by triturating and filtrating in ethyl ether.

General Procedure C

A solution of the corresponding Boc-aminoacid-Lamellarin in a 3.0M solution of HCl in ethyl acetate was stirred at room temperature for 30 min. The resulting suspension was filtered and the solid was washed with ethyl acetate and hexanes to provide the corresponding Lamellarin.

General Procedure D

To a 0.01M suspension of Lamellarin (1 eq.) in anhydrous dichloromethane, the corresponding carboxylic acid (2 eq. per hydroxy group), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2 eq. per hydroxy group) and dimethyl-aminopyridine (0.2 eq. per hydroxy group) were added. The mixture was stirred under argon atmosphere at room temperature for 6 h. The resulting solution was diluted with dichloromethane, washed with water and saturated sodium bicarbonate. The organic layer was dried over anhydrous sodium sulfate and the solvent removed under vacuum. The residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure E

A 0.02M solution of the corresponding Lamellarin (1 eq.) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (1.3 eq.) in chloroform was heated at 65° C. until the reaction was completed. The mixture was cooled to room temperature and the solvent evaporated under reduced pressure. The residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure F

To a 0.01M solution of Lamellarin (1 eq.) in dichloromethane, pyridine (1.1 eq. per hydroxy group) and the corresponding acid chloride (1.1 eq. per hydroxy group) were added under argon atmosphere and stirred at room temperature for 3 h. The reaction mixture was washed with saturated solution of sodium bicarbonate, dried over anhydrous sodium sulfate, filtered and the solvent was evaporated under reduced pressure. The residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure G

Iodo-acetic acid 5-isopropoxy-2-(4-isopropoxy-3-methoxy-phenylethynyl)-4-methoxy-phenyl ester (1 eq.) was added in one portion to a 0.1M solution of the corresponding dihydro-isoquinoline or isoquinoline (1.1 eq.) in anhydrous dimethylacetamide under argon atmosphere. The solution was stirred at room temperature for 14 hours, then triethylamine (1.1 eq.) was added and the reaction mixture was heated at 80° C. for 19 hours. The mixture was cooled, Fremy's Salt (1.1 eq) and sodium carbonate saturated solution was added and the suspension was stirred for 1 hour. The mixture was treated with sodium bicarbonate saturated solution and extracted with dichloromethane. The organic layers were dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. The resulted residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure H

Iodo-acetic acid 5-isopropoxy-2-(4-isopropoxy-3-methoxyphenyl-ethynyl)-4-methoxy-phenyl ester (1 eq.) was added in one portion to a 0.1 M solution of the corresponding dihydro-isoquinoline or isoquinoline (1.1 eq.) in dry 1,2-dichloroethane under argon atmosphere. The solution was stirred at room temperature for 14 hours, then diisopropylethylamine (1.1 eq.) was added and the reaction mixture was heated at 85° C. for 32 hours. The resulting mixture was cooled, silica gel (1 g per mmol) was added and the solvent was evaporated under reduced pressure. The resulted residue was subjected to flash chromatography on silica gel (sequential elution with 5:5:1 to 5:5:2 hexane-dichloromethane-ether) to provide the corresponding Lamellarin.

General Procedure I

To a 0.015M suspension of the corresponding Lamellarin (1 eq.) in anhydrous dichloromethane at 0° C., N-phenyltrifluoro-methanesulfonimide (4 eq.), triethylamine (7 eq.) and dimethyl amino-pyridine (0.2 eq.) were added and the mixture was stirred to room temperature for 3 h. The mixture was diluted with dichloromethane, washed with sodium bicarbonate saturated solution, dried over anhydrous sodium sulfate and evaporated to dryness. The residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure J

To a 0.005M solution of Lamellarin (1 eq.) in methanol, palladium/C 10% (1 eq., w/w) was added and the resulting suspension was stirred at room temperature under hydrogen atmosphere. The mixture was filtered on celite and washed with dichloromethane. Evaporation of the solvent gave the corresponding Lamellarin.

General Procedure K

To a 0.01M suspension of Lamellarin (1 eq.) in anhydrous dichloromethane, the corresponding carboxylic acid (2 eq. per hydroxy group), 1,3-dicyclohexylcarbodiimide (2 eq. per hydroxy group) and dimethyl-aminopyridine (0.2 eq. per hydroxy group) were added. The mixture was stirred under argon atmosphere at room temperature for 6 h. The resulting solution was diluted with dichloromethane, washed with water and saturated sodium bicarbonate. The organic layer was dried over anhydrous sodium sulfate and the solvent removed under vacuum. The residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General Procedure L

A 0.01M mixture of the corresponding Lamellarin in acetic anhydride/pyridine (1:2) was stirred overnight at room temperature under argon atmosphere. The solvent was evaporated under reduced pressure to provide the acylated-Lamellarin.

General Procedure M

To a 0.03M solution of 189 (1 eq.) in toluene/ethanol (10:1) under argon atmosphere, the corresponding boronic acid (2 eq.), tetrakis-triphenylphosphine palladium(0) (0.05 eq.) and sodium carbonate 2M (6 eq.) were added. The resulting mixture was heated at 90° C. for 16 hours, then water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with sodium hydroxide 1M, water and brine. After drying over sodium sulfate and evaporation of the solvent under reduced pressure, the residue was purified by chromatography on silica gel to provide the corresponding Lamellarin.

General procedure A (starting from 104) and chromatography on silica gel (CH₂Cl₂:MeOH, from 20:1 to 15:1) to afford 1 (2.27 g, 95%).

¹H NMR (300 MHz, CDCl₃) δ 7.13 (d, J=7.8 Hz, 1H), 7.06 (dd, J=7.8, 1.7 Hz, 1H), 6.98 (d, J=1.7 Hz, 1H), 6.93 (s, 1H), 6.59 (s, 1H), 6.38 (s, 1H), 6.02 (s, 1H), 5.85 (s, 1H), 5.82 (s, 1H), 5.00-4.80 (m, 1H), 4.70-4.50 (m, 1H), 3.88 (s, 3H), 3.87 (s, 3H), 3.49 (s, 3H), 3.36 (s, 3H), 3.20-3.00 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 150.3, 147.2, 146.3, 146.1, 145.5, 145.4, 143.3, 135.4, 135.3, 128.1, 127.3, 124.2, 123.2, 115.5, 115.1, 113.8, 113.4, 113.0, 110.1, 103.9, 103.3, 101.8, 61.0, 56.2, 55.6, 55.5, 42.0, 21.4.

MS (ESI) m/z: 532 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂:MeOH, 20:1).

General procedure A (starting from 27) and chromatography on silica gel (CH₂Cl₂:MeOH, from 20:1 to 10:1) to afford 2 (8 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 9.18 (d, J=7.5 Hz, 1H), 7.40 (d, J=7.5 Hz, 1H), 7.20-7.16 (m, 2H), 7.10 (s, 1H), 6.99 (s, 1H), 6.80 (s, 1H), 6.68 (s, 1H), 6.20 (br s, 1H), 5.80 (br s, 2H), 5.80 (br s, 2H), 3.94 (s, 3H), 3.91 (s, 3H), 3.52 (s, 3H), 3.46 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 155.5, 151.9, 147.3, 147.0, 146.3, 145.7, 144.5, 143.3, 134.8, 133.7, 129.3, 127.5, 124.7, 122.3, 121.4, 119.7, 115.2, 113.9, 113.8, 111.9, 109.8, 106.9, 104.6, 103.5, 98.3, 61.2, 56.3, 55.6, 55.1.

MS (ESI) m/z: 529 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂:MeOH, 20:1).

General procedure A (starting from 107) and chromatography on silica gel (EtOAc, 100%) to afford 3 (92 mg, 43%).

¹H NMR (300 MHz, DMSO-d₆) δ 9.92 (s, 1H), 9.81 (s, 1H), 9.32 (s, 1H), 8.98 (d, J=7.3 Hz, 1H), 7.22-6.98 (m, 6H), 6.85 (s, 1H), 6.70 (s, 1H), 3.75 (s, 3H), 3.36 (s, 6H).

¹³C NMR (75 MHz, DMSO-d₆) δ 154.3, 148.7, 148.5, 148.3, 147.8, 146.8, 146.3, 144.6, 134.1, 129.2, 128.9, 125.5, 124.7, 123.9, 117.6, 116.4, 115.1, 113.9, 112.3, 111.5, 110.8, 106.4, 105.7, 105.4, 103.7, 56.0, 55.1, 54.5. MS (APCI) m/z: 500 (M+1)⁺.

Rf: 0.60 (EtOAc).

General procedure A (starting from 50) and chromatography on silica gel (CH₂Cl₂:MeOH, 10:1) to provide 4 (9.1 mg, 76%).

¹H NMR (300 MHz, CD₃OD) δ 7.07-7.05 (m, 2H), 6.99-6.97 (m, 1H), 6.80 (br s, 2H), 6.75 (s, 1H), 6.71 (s, 1H), 4.75 (m, 1H), 3.82 (s, 3H), 3.43 (s, 3H), 3.36 (s, 3H), 3.02 (br t, 2H).

MS (ESI) m/z: 501 (M+1)⁺.

Rf: 0.32 (CH₂Cl₂:MeOH, 10:1).

General procedure B (starting from 6) to afford 5 (30 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.07 (br s, 1H), 7.60-7.10 (m, 5H), 6.90-6.60 (m, 2H), 4.85-4.60 (m, 3H), 4.10-3.90 (m, 3H), 3.84 (s, 6H), 3.50 (s, 3H), 3.44 (s, 3H), 1.80-1.50 (m, 18H).

MS (ESI) m/z: 956 (M+1)⁺.

General procedure D (starting from 2 and Boc-Ala-Ala-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 6 (56 mg, 87%).

¹H NMR (300 MHz, CDCl₃) δ 9.02 (br s, 1H), 7.40-7.10 (m, 4H), 7.10-6.90 (m, 3H), 6.90-6.60 (m, 3H), 5.30-5.00 (m, 3H), 5.00-4.75 (m, 3H), 4.26 (br s, 3H), 3.85 (s, 6H), 3.47 (s, 3H), 3.43 (s, 3H), 1.80-1.30 (m, 45H).

¹³C NMR (75 MHz, CDCl₃) δ 173.1, 172.6, 171.3, 171.2, 171.1, 171.0, 170.9, 155.8, 154.8, 153.4, 153.3, 153.2, 152.4, 152.3, 147.6, 146.8, 145.5, 143.9, 141.8, 140.2, 140.0, 139.5, 138.9, 135.2, 134.8, 133.3, 133.2, 128.3, 128.2, 124.1, 123.8, 123.6, 121.1, 118.4, 115.9, 115.7, 115.5, 112.2, 111.9, 111.4, 109.1, 106.8, 106.4, 106.1, 104.5, 103.7, 80.5, 61.1, 56.6, 55.9, 55.8, 53.6, 50.2, 48.6, 48.4, 48.3, 28.5, 18.6, 18.3.

MS (ESI) m/z: 1279 (M+23)⁺.

Rf: 0.24 (CH₂Cl₂:MeOH, 20:1).

General procedure D (starting from 109 and Boc-Ala-Ala-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 7 (46 mg, 75%).

¹H NMR (300 MHz, CDCl₃) δ 7.26-6.50 (m, 10H), 5.13-5.11 (m, 3H), 4.87-4.65 (m, 5H), 4.23 (br s, 3H), 3.78 (s, 3H), 3.39 (s, 3H), 3.32 (s, 3H), 3.06 (br t, 2H), 1.63-1.53 (m, 9H), 1.44-1.35 (m, 36H).

MS (ESI) m/z: 1250 (M+23)⁺.

Rf: 0.40 (CH₂Cl₂:MeOH, 20:1).

General procedure E (starting from 11, reaction time 21 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 8 (16 mg, 70%).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.3 Hz, 1H), 7.39-7.20 (m, 20H), 7.08-7.03 (m, 2H), 6.80 (s, 1H), 3.79 (s, 3H), 3.42 (s, 6H), 3.16-3.06 (m, 6H), 3.00-2.90 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 170.8, 170.7, 170.6, 155.1, 152.4, 151.0, 147.7, 145.4, 140.9, 140.2, 140.1 (2C), 139.7, 134.2, 133.5, 128.5 (6C), 128.4 (2C), 128.4 (4C), 128.2, 126.4, 126.4 (2C), 124.0, 123.8, 123.6, 123.6, 123.1, 120.7, 115.6, 115.0, 112.8, 112.3, 112.1, 109.0, 106.4, 106.1, 56.2, 55.7, 55.6, 35.4 (3C), 30.9, 30.8 (2C).

MS (APCI) m/z: 896 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂:MeOH, 200:1).

General procedure B (starting from 10) to afford 9 (12 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.15 (d, J=7.5 Hz, 1H), 7.60-7.50 (m, 2H), 7.30-7.35 (m, 2H), 7.29 (s, 1H), 7.23 (s, 1H), 6.89 (s, 1H), 4.73 (br t, 1H), 4.54 (br t, 1H), 4.44 (br t, 1H), 4.30-4.10 (m, 6H), 3.89 (s, 6H), 3.54 (s, 3H), 3.47 (s, 3H).

MS (ESI) m/z: 791 (M+1)⁺.

General procedure E (starting from 60, reaction time 22 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 10 (17 mg, 61%).

¹H NMR (300 MHz, CDCl₃) δ 9.05 (br s, 1H), 7.40-7.00 (m, 6H), 6.72 (d, J=12.4 Hz, 1H), 5.70 (br s, 1H), 5.55 (br s, 2H), 4.90-4.60 (m, 3H), 4.32 (br s, 2H), 4.20-3.80 (m, 9H), 3.49 (s, 3H), 3.46 (s, 3H), 3.00-2.50 (m, 3H), 1.51 (s, 18H), 1.47 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 169.6, 169.2, 155.6, 154.5, 153.0, 151.7, 146.8, 145.5, 141.3, 139.8, 139.2, 138.7, 134.7, 133.0, 127.9, 124.0, 123.4, 120.9, 118.2, 115.8, 115.3, 112.1, 111.8, 108.9, 106.9, 106.2, 104.3, 80.6, 64.0, 63.7, 61.1, 56.5, 56.1, 55.8, 55.6, 28.3.

MS (ESI) m/z: 1113 (M+23)⁺, 1091 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂:MeOH, 20:1).

General procedure F (starting from 109 and hydrocinnamoyl chloride) and chromatography on silica gel (CH₂Cl₂:MeOH, from 200:1 to 100:1) to afford 11 (31 mg, 69%).

¹H NMR (300 MHz, CDCl₃) δ 7.37-7.21 (m, 15H), 7.13-7.02 (m, 4H), 6.87 (s, 1H), 6.77 (s, 1H), 6.68 (s, 1H), 4.92-4.83 (m, 1H), 4.79-4.70 (m, 1H), 3.76 (s, 3H), 3.39 (s, 3H), 3.32 (s, 3H), 3.13-3.04 (m, 8H), 2.97-2.87 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 171.0, 170.7, 170.6, 155.1, 152.2, 149.8, 147.7, 144.9, 140.1 (3C), 140.0, 139.4, 138.9, 135.0, 133.9, 128.5 (6C), 128.4 (6C), 127.1, 126.4, 126.4, 126.4, 125.9, 125.6, 123.8, 123.1, 122.6, 116.0, 115.9, 114.9, 114.6, 111.9, 109.7, 105.4, 56.1, 55.7, 55.5, 42.4, 35.5 (3C), 30.9, 30.9, 30.8, 28.0.

MS (ESI) m/z: 898 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂:MeOH, 200:1).

General procedure E (starting from 106, reaction time 17 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 50:1) to affrord 12 (21 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.3 Hz, 1H), 7.38 (s, 1H), 7.29-7.13 (m, 5H), 7.06 (d, J=7.5 Hz, 1H), 6.81 (s, 1H), 3.82 (s, 3H), 3.45 (s, 6H), 2.67-2.57 (m, 6H), 1.82 (m, 21H), 1.40-1.13 (m, 12H), 1.04-0.85 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 172.0, 171.9 (2C), 155.1, 152.5, 151.1, 147.9, 145.5, 141.0, 140.4, 139.9, 134.1, 133.6, 128.3, 124.1, 123.8, 123.6, 123.1, 120.7, 115.6, 115.0, 112.8, 112.3, 112.2, 109.0, 106.4, 106.1, 56.2, 55.8, 55.7, 37.3 (3C), 37.1 (3C), 33.0 (6C), 32.3, 32.3, 32.2, 31.6 (3C), 26.5 (2C), 26.3 (2C), 26.3 (2C).

MS (ESI) m/z: 914 (M+1)⁺.

Rf: 0.17 (hexane:EtOAc, 4:1).

General procedure B (starting from 58) to afford 13 (10.5 mg, quant).

¹H NMR (300 MHz, CD₃OD) δ 8.93 (d, J=7.1 Hz, 1H), 7.58 (d, J=7.8 Hz, 1H), 7.47-7.34 (m, 12H), 7.20 (s, 1H), 7.12 (d, J=7.3 Hz, 1H), 7.08 (s, 1H), 7.05 (d, J=4.0 Hz, 1H), 6.83 (s, 1H), 4.76 (br t, J=6.8 Hz, 1H), 4.61 (m, 1H), 3.95 (s, 3H), 3.86 (s, 3H), 3.46-3.31 (m, 7H).

MS (ESI) m/z: 830.1 (M+23)⁺, 808 (M+1)⁺.

General procedure B (starting from 15) to afford 14 (11.6 mg, quant).

¹H NMR (300 MHz, CD₃OD) δ 9.05 (m, 1H), 7.58-7.47 (m, 2H), 7.38-7.31 (m, 3H), 7.38-7.10 (m, 4H), 6.88 (br d, 1H), 4.36 (m, 1H), 4.25 (m, 2H), 3.92 (s, 3H), 3.88 (s, 3H), 3.47 (s, 6H), 2.50-2.48 (m, 2H), 1.27 (d, J=6.1 Hz, 6H), 1.20 (d, J=6.8 Hz, 6H).

MS (ESI) m/z: 712 (M+1)⁺.

General procedure E (starting from 65, reaction time 20 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 15 (29.0 mg, 90%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.6 Hz, 1H), 7.32-7.23 (m, 3H), 7.12-7.05 (m, 4H), 6.80 (d, J=9.2 Hz, 1H), 5.09 (br d, 2H), 4.52 (br s, 2H), 3.98 (s, 3H), 3.80 (s, 3H), 3.50 (s, 3H), 3.43 (s, 3H), 2.43-2.37 (m, 2H), 1.49 (s, 9H), 1.46 (s, 9H), 1.14-0.99 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4, 155.7 (2C), 154.9, 152.2, 150.3, 149.6, 147.5, 145.5, 139.9, 139.3, 134.8, 134.2, 128.3, 128.2, 124.7, 123.9 (2C), 123.1, 118.9, 116.0, 115.3, 113.0, 112.1, 110.9, 108.4, 107.4, 106.2, 105.1, 80.0 (2C), 58.6 (2C), 56.0, 55.9, 55.7, 55.6, 31.3, 31.2, 28.3 (9C), 19.2, 19.0, 17.1 (2C).

MS (ESI) m/z: 934.2 (M+23)⁺, 912 (M+1)⁺.

Rf: 0.54 (CH₂Cl₂:MeOH, 60:1).

General procedure B (starting from 97) to afford 16 (31 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.11 (dd, J=7.5, 2.3 Hz, 1H), 7.60-7.50 (m, 2H), 7.35 (t, J=6.6 Hz, 1H), 7.25-7.20 (m, 3H), 6.86 (d, J=9.5 Hz, 1H), 4.66 (q, J=7.3 Hz, 1H), 4.52 (q, J=7.3 Hz, 1H), 4.38 (q, J=7.1 Hz, 1H), 3.90 (d, J=3.2 Hz, 3H), 3.89 (d, J=1.8 Hz, 3H), 3.53 (d, J=2.7 Hz, 3H), 3.47 (s, 3H), 1.85 (d, J=7.0 Hz, 3H), 1.80 (d, J=7.1 Hz, 3H), 1.69 (dd, J=7.1, 4.0 Hz, 3H).

MS (ESI) m/z: 743 (M+1)⁺.

General procedure B (starting from 122) to afford 17 (21 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.13-9.09 (m, 1H), 7.63-7.52 (m, 3H), 7.44-7.22 (m, 4H), 6.89 (d, J=9.2 Hz, 1H), 4.36 (d, J=4.4 Hz, 1H), 4.32 (d, J=4.0 Hz, 1H), 4.25 (t, J=3.8 Hz, 1H), 3.91 (s, 3H), 3.48 (s, 6H), 2.61-2.44 (m, 3H), 1.29-1.19 (m, 18H).

MS (ESI) m/z: 797 (M+1)⁺.

General procedure B (starting from 84) to afford 18 (21.6 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.44-7.37 (m, 11H), 7.23-7.20 (m, 2H), 7.03 (s, 1H), 6.90 (s, 1H), 6.78 (m, 1H), 6.67 (s, 1H), 4.72-4.60 (m, 4H), 3.90 (s, 3H), 3.81 (s, 3H), 3.44 (s, 3H), 3.39-3.30 (m, 4H), 3.11 (br t, 2H).

MS (ESI) m/z: 810 (M+1)⁺.

General procedure B (starting from 65) to afford 19 (43.3 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.43-7.40 (m, 2H), 7.25-7.17 (m, 3H), 6.94 (br s, 1H), 6.80 (d, J=13.4 Hz, 1H), 6.69 (d, J=9.03 Hz, 1H), 4.90 (m, 2H), 4.32 (m, 1H), 4.22 (m, 2H), 3.86 (s, 3H), 3.45 (s, 3H), 3.36 (s, 3H), 3.13 (br t, 2H), 2.58-2.40 (m, 2H), 1.25 (br d, 6H), 1.17 (br d, 6H).

MS (ESI) m/z: 736 (M+23)⁺, 714 (M+1)⁺.

General procedure B (starting from 77) to afford 20 (11.7 mg, quant).

¹H NMR (300 MHz, CD₃OD) δ 9.03 (d, J=7.3 Hz, 1H), 7.57-7.48 (m, 2H), 7.37-7.32 (m, 1H), 7.26-7.25 (m, 1H), 7.21-7.20 (m, 2H), 7.11 (d, J=6.8 Hz, 1H), 6.87 (d, J=8.3 Hz, 1H), 4.52 (br q, J=6.8 Hz, 1H), 4.42 (br q, J=6.8 Hz, 1H), 3.91 (s, 3H), 3.90 (s, 3H), 3.48 (s, 6H), 1.85 (d, J=7.3 Hz, 1H), 1.71 (d, J=7.1 Hz, 1H).

MS (ESI) m/z: 678 (M+23)⁺, 656 (M+1)⁺.

General procedure D (starting from 95 and Boc-Ala-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 21 (83.2 mg, 100%).

¹H NMR (300 MHz, CDCl₃) δ 7.23 (br s, 1H), 7.16-7.10 (m, 3H), 6.76 (s, 1H), 6.71-6.56 (m, 2H), 5.10 (m, 1H), 4.92-4.70 (m, 2H), 4.60-4.58 (m, 2H), 3.89 (s, 3H), 3.78 (s, 3H), 3.41 (s, 3H), 3.40 (s, 3H), 3.13 (t, J=7.1 Hz, 2H), 1.63-1.46 (m, 24H).

¹³C NMR (75 MHz, CDCl₃) δ 171.3, 154.9, 151.9, 149.1, 147.6, 147.3, 144.8, 139.6, 138.4, 135.8, 134.4, 127.0, 126.4, 123.6, 123.3, 119.5, 116.2, 114.8, 114.6, 114.3, 111.6, 110.9, 108.4, 105.4, 79.9 (2C), 56.1, 55.8, 55.7, 55.3, 49.3 (2C), 42.4, 28.4, 28.2 (6C), 18.5 (2C).

MS (ESI) m/z: 880 (M+23)⁺, 857 (M+1)⁺.

Rf: 0.15 (CH₂Cl₂:MeOH, 60:1).

To a suspension of 109 (50 mg, 0.0997 mmol) in anhydrous CH₂Cl₂ (2 mL) under Argon at 0° C., Et₃N (83 μL, 0.5982 mmol) and methanesulfonyl chloride (47 μL, 0.5982 mmol) were added. The resulting mixture was stirred at 23° C. for 6 h, then quenched with H₂O and extracted with CH₂Cl₂ (3×20 mL).

The combined organic layers were washed with saturated aqueous solution of NaHCO₃, dried over anhydrous Na₂SO₄, filtered, and evaporated under vacuum. The resulting residue was purified on silica gel (CH₂Cl₂:MeOH, 80:1) to afford 22 as a pale yellow solid (47 mg, 64%).

¹H NMR (300 MHz, CDCl₃) δ 7.52 (d, J=8.1 Hz, 1H), 7.30 (s, 1H), 7.23-7.20 (m, 2H), 7.17 (d, J=1.6 Hz, 1H), 6.75 (s, 1H), 6.65 (s, 1H), 4.99-4.90 (m, 1H), 4.71-4.61 (m, 1H), 3.92 (s, 3H), 3.46 (s, 3H), 3.38 (s, 3H), 3.34 (s, 3H), 3.19 (s, 3H), 3.18 (s, 3H), 3.14 (t, J=6.0 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 154.5, 152.8, 150.1, 148.0, 144.6, 138.0, 137.7, 136.9, 135.4, 134.4, 126.5, 126.4, 126.3, 125.6, 124.4, 123.3, 117.0, 115.8, 115.4, 115.2, 113.5, 109.9, 105.6.

MS (ESI) m/z: 736 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 80:1).

General procedure B (starting from 114) to give 23 (20 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.90-7.60 (m, 12H), 7.45-7.20 (m, 16H), 6.80-6.70 (m, 2H), 4.80-4.40 (m, 5H), 4.35-4.20 (m, 3H), 3.74 (d, J=2.9 Hz, 3H), 3.71 (d, J=2.3 Hz, 3H), 3.55-3.30 (m, 12H) 3.35-3.00 (m, 6H), 2.91 (br s, 2H).

MS (ESI) m/z: 1375 (M)⁺.

General procedure B (starting from 29) to afford 24 (27 mg, quant.)

¹H NMR (300 MHz, CD₃OD) δ 7.68 (d, J=8.1 Hz, 1H), 7.62 (s, 1H), 7.60 (s, 1H), 7.44 (s, 1H), 7.42 (s, 3H), 7.34 (s, 1H), 7.30 (s, 1H), 7.29 (s, 1H), 7.21-7.16 (m, 5H), 7.12-7.09 (m, 3H), 6.98 (s, 1H), 6.85 (d, J=2.0 Hz, 1H), 6.77-6.76 (m, 2H), 4.73-4.69 (m, 3H), 4.60 (br t, 2H), 3.87 (s, 3H), 3.77-3.34 (m, 6H), 3.43 (s, 3H), 3.34 (s, 3H).

MS (ESI) m/z: 1060 (M+1)⁺.

General procedure B (starting from 113) to afford 25 (20 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.08 (d, J=7.5 Hz, 1H), 7.60 (d, J=2.2 Hz, 1H), 7.53 (s, 1H), 7.49-7.36 (m, 17H), 7.30 (d, J=3.5 Hz, 1H), 7.20 (d, J=7.5 Hz, 1H), 7.08 (d, J=4.8 Hz, 1H), 6.87 (d, J=4.2 Hz, 1H), 4.76 (t, J=7.0 Hz, 1H), 4.70 (t, J=6.9 Hz, 1H), 4.63 (t, J=6.2 Hz, 1H), 3.95 (s, 3H), 3.47 (s, 6H), 3.61-3.36 (m, 6H).

MS (ESI) m/z: 941 (M+1)⁺.

General procedure A (starting from 111) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 26 (116 mg, 82%).

¹H NMR (300 MHz, CDCl₃) δ 9.19 (d, J=7.3 Hz, 1H), 7.19-6.98 (m, 7H), 6.71 (s, 1H), 5.86-5.85 (br s, 2H), 3.97 (s, 3H), 3.90 (s, 3H), 3.52 (s, 3H), 3.48 (s, 3H).

¹³C NMR (75 MHz, DMSO-d₆) δ 154.2, 149.7, 148.7, 148.6, 147.7, 146.8, 146.3, 144.4, 133.5, 128.7, 125.5, 124.2, 124.0, 121.9, 118.2, 116.2, 115.2, 112.2, 110.8, 108.3, 107.7, 106.5, 105.6, 104.8, 103.6, 56.0, 55.4, 55.0, 54.4.

MS (ESI) m/z: 536 (M+23)⁺, 514 (M+1)⁺.

Rf: 0.45 (CH₂Cl₂:MeOH, 20:1).

General procedure G (starting from 6,7-dimethoxy-5-isopropoxyisoquinoline) and chromatography on silica gel (hexane:EtOAc, from 3:1 to 2:1) to afford 27 (15 mg, 7%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.5 Hz, 1H), 7.43 (d, J=7.6 Hz, 1H), 7.20-7.15 (m, 3H), 7.01 (s, 1H), 6.97 (s, 1H), 6.72 (s, 1H), 4.75-4.50 (m, 3H), 4.65-4.50 (m, 3H), 3.90 (s, 3H), 3.85 (s, 3H), 3.44 (s, 3H), 1.50-1.35 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 159.3, 155.6, 153.2, 151.4, 147.8, 147.2, 146.6, 146.5, 146.4, 142.5, 133.8, 129.3, 128.7, 123.8, 122.6, 121.3, 120.8, 116.9, 114.9, 111.9, 109.9, 107.7, 105.4, 103.4, 101.4, 76.4, 71.8, 71.4, 60.7, 56.2, 55.4, 55.1, 22.7, 21.9, 21.8.

MS (ESI) m/z: 656 (M+1)⁺.

Rf: 0.20 (hexane:EtOAc, 2:1).

General procedure D (starting from 109 and n-octanoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 28 (42 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 7.21-7.07 (m, 4H), 6.94 (s, 1H), 6.79 (s, 1H), 6.70 (s, 1H), 4.93-4.84 (m, 1H), 4.79-4.70 (m, 1H), 3.80 (s, 3H), 3.42 (s, 3H), 3.35 (s, 3H), 3.11 (t, J=6.6 Hz, 2H), 2.63-2.53 (m, 6H), 1.83-1.69 (m, 6H), 1.41-1.30 (m, 24H), 0.93-0.87 (m, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 171.6, 171.5, 155.1, 152.3, 149.9, 147.7, 144.9, 140.1, 139.5, 139.1, 135.1, 133.8, 127.1, 125.9, 125.5, 123.9, 123.1, 122.6, 115.9, 114.9, 114.6, 111.9, 109.7, 105.4, 56.1, 55.7, 55.5, 42.4, 34.0 (3C), 31.7 (3C), 29.0 (2C), 28.9 (4C), 28.0, 25.0 (2C), 24.9, 22.6 (3C), 14.0 (3C).

MS (ESI) m/z: 902 (M+23)⁺, 880 (M+1)⁺.

Rf: 0.31 (CH₂Cl₂:MeOH, 100:1).

General procedure D (starting from 109 and Boc-L-Trp-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, from 30:1 to 15:1) to afford 29 (115 mg, 85%).

¹H NMR (300 MHz, CDCl₃) δ 8.35 (s, 1H), 8.28 (s, 2H), 7.68-7-62 (m, 3H), 7.39-7.36 (m, 3H), 7.26-7.07 (m, 12H), 6.90 (s, 1H), 6.72 (s, 1H), 6.65 (br s, 2H), 5.15-5.12 (m, 2H), 5.00-4.59 (m, 6H), 3.75 (s, 3H), 3.52-3.28 (m, 12H), 3.00 (br t, 2H), 1.43 (s, 27H).

¹³C NMR (75 MHz, CDCl₃) δ 170.7, 170.4, 170.4, 155.3 (2C), 154.9, 152.0, 149.6, 147.5, 144.6, 139.6, 139.0, 138.4, 136.1 (3C), 134.9, 134.0, 127.7 (3C), 126.8, 125.9, 125.5, 123.8, 123.1 (3C), 122.5, 122.0 (3C), 119.5 (3C), 118.6 (3C), 116.0, 115.8, 114.7, 111.7, 111.3 (3C), 109.5 (3C), 105.3, 80.0 (3C), 56.0 (2C), 55.6, 55.4, 54.4 (2C), 42.3, 28.2 (12C), 27.7.

MS (ESI) m/z: 1382 (M+23)⁺.

Rf: 0.13 (CH₂Cl₂:MeOH, 30:1).

General procedure B (starting from 117) to afford 30 (11 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.15 (d, J=7.6 Hz, 1H), 7.65-7.55 (m, 2H), 7.50-7.20 (m, 4H), 6.87 (d, J=12.3 Hz, 1H), 4.90-4.70 (m, 3H), 3.90 (s, 6H), 3.85-3.40 (m, 12H), 2.90-2.00 (m, 12H).

MS (ESI) m/z: 821 (M+1)⁺.

General procedure B (starting from 120) to afford 31 (31 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.09 (d, J=7.8 Hz, 1H), 7.80-7.40 (m, 18H), 7.30-7.00 (m, 3H), 6.87 (d, J=5.3 Hz, 1H), 4.80-4.60 (m, 3H), 3.92 (s, 3H), 3.90 (s, 3H), 3.80-3.40 (m, 12H).

MS (ESI) m/z: 971 (M)⁺.

General procedure B (starting from 34) to afford 32 (19 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.46-7.44 (m, 2H), 7.29-7.25 (m, 1H), 7.17-7.14 (m, 2H), 6.90-6.78 (m, 2H), 4.76 (br t, 2H), 4.33-4.21 (m, 3H), 3.88 (s, 3H), 3.45 (s, 3H), 3.38 (s, 3H), 3.16 (br t, 2H), 2.59-2.43 (m, 3H), 1.27-1.10 (m, 18H).

MS (ESI) m/z: 799 (M+1)⁺.

General procedure B (starting from 127) to afford 33 (19 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 8.96-8.90 (m, 1H), 7.67-7.52 (m, 3H), 7.40-7.24 (m, 2H), 7.13-7.08 (m, 2H), 6.84-6.80 (m, 1H), 4.86-4.67 (m, 3H), 3.95 (s, 3H), 3.55-3.43 (m, 12H), 2.66-2.35 (m, 6H), 2.27-2.14 (m, 6H).

MS (ESI) m/z: 791 (M+1)⁺.

General procedure D (starting from 109 and Boc-D-Val-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 50:1) to afford 34 (100 mg, 91%).

¹H NMR (300 MHz, CDCl₃) δ 7.26-7.08 (m, 4H), 6.97 (s, 1H), 6.77 (d, J=7.1 Hz, 1H), 6.69 (d, J=9.3 Hz, 1H), 5.07-5.05 (m, 3H), 4.96-4.90 (m, 1H), 4.75-4.70 (m, 1H), 4.55-4.47 (m, 3H), 3.78 (s, 3H), 3.40 (s, 3H), 3.33 (s, 3H), 3.21 (br t, 2H), 2.45-2.30 (m, 3H), 1.49 (s, 9H), 1.47 (s, 18H), 1.12-0.99 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.5, 170.3 (2C), 155.6, 154.9, 152.0, 149.7, 147.5, 144.8, 139.6, 139.1, 138.5, 134.9, 134.2, 126.9, 125.9, 125.7, 123.8, 123.1, 122.5, 116.1, 115.8, 114.9, 114.6, 111.8, 109.6, 105.4, 79.9 (3C), 58.5, 55.9, 55.5, 55.5, 55.3, 55.2, 42.3, 31.5, 31.2, 31.1, 28.2 (9C), 27.9, 19.0 (2C), 17.1 (4C).

MS (ESI) m/z: 1099 (M+1)⁺.

Rf: 0.35 (CH₂Cl₂:MeOH 50:1).

General procedure B (starting from 129) to afford 35 (13 mg, 98%).

¹H NMR (300 MHz, CD₃OD) δ 9.14 (dd, J=7.5, 3.0 Hz, 1H), 7.60-7.50 (m, 2H), 7.50-7.20 (m, 4H), 6.87 (d, J=11.1 Hz, 1H), 4.60-4.50 (m, 1H), 4.40-4.30 (m, 1H), 4.25-4.20 (m, 1H), 3.89 (s, 3H), 3.88 (s, 3H), 3.52 (s, 3H), 3.47 (s, 3H), 2.80-2.40 (m, 3H), 1.40-1.10 (m, 18H).

MS (ESI) m/z: 827 (M+1)⁺.

General procedure B (starting from 38) to afford 36 (21 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.09-9.04 (m, 1H), 7.62-7.51 (m, 3H), 7.41-7.32 (m, 2H), 7.23-7.18 (m, 2H), 6.88 (d, J=9.0 Hz, 1H), 4.36 (d, J=4.4 Hz, 1H), 4.31 (d, J=4.4 Hz, 1H), 4.24 (t, J=4.4 Hz, 1H), 3.92 (s, 3H), 3.48 (s, 6H), 2.62-2.43 (m, 3H), 1.29-1.19 (m, 18H).

MS (APCI) m/z: 797 (M+1)⁺.

General procedure C (starting from 144) to afford 37 as a white solid (654 mg, 83%).

¹H NMR (300 MHz, CD₃OD) δ 7.46-7.43 (m, 2H), 7.26-7.16 (m, 3H), 6.89-6.78 (m, 2H), 4.80 (br t, 2H), 4.33 (s, 1H), 4.24 (s, 2H), 3.84 (s, 3H), 3.45 (s, 3H), 3.37 (s, 3H), 3.18 (br t, 2H), 2.60-2.40 (m, 3H), 1.27-1.17 (m, 18H).

¹³C NMR (75 MHz, CD₃OD) δ 168.7, 168.3 (2C), 156.2, 153.5, 150.8, 148.8, 146.1, 140.6, 140.0, 139.3, 136.6, 136.4, 128.4, 128.0, 127.5, 125.2, 124.7, 123.7, 117.9, 117.6, 116.5, 116.2, 112.8, 110.9, 106.7, 59.5, 59.4, 56.9, 56.4, 56.2, 56.0, 43.7, 31.3 (3C), 28.8, 18.3 (2C), 18.2 (4C).

MS (ESI) m/z: 799 (M+1)⁺.

General procedure E (starting from 144, overnight) and chromatography on silica gel (CH₂Cl₂:MeOH, from 100:1 to 50:1) to afford 38 (43 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.3 Hz, 1H), 7.40 (s, 1H), 7.30-7.13 (m, 5H), 7.04 (d, J=7.3 Hz, 1H), 6.79 (d, J=7.5 Hz, 1H), 5.10-5.06 (m, 3H), 4.56-4.48 (m, 3H), 3.81 (s, 3H), 3.43 (s, 6H), 2.45-2.32 (m, 3H), 1.49 (s, 9H), 1.47 (s, 18H), 1.14-1.00 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4 (3C), 155.7, 155.0, 152.3, 150.9, 147.6, 145.4, 140.6, 140.0, 139.4, 134.5, 133.5, 130.9, 128.8, 128.2, 128.1, 124.1, 123.8, 123.6, 123.2, 120.8, 115.9, 115.1, 112.8, 112.3, 112.2, 109.1, 106.4, 106.2, 80.0 (3C), 58.5, 56.0, 55.6, 55.6, 55.5, 55.5, 31.3 (2C), 31.2, 28.3 (9C), 19.2, 19.1, 17.2 (2C), 17.1 (2C).

MS (ESI) m/z: 1119 (M+23)⁺, 1097 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 100:1).

General procedure B (starting from 146) to afford 39 (19 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.15-9.12 (m, 1H), 7.64 (d, J=2.7 Hz, 1H), 7.58-7.52 (m, 2H), 7.40-7.29 (m, 2H), 7.26-7.22 (m, 2H), 6.89 (d, J=7.3 Hz, 1H), 4.45-4.31 (m, 3H), 3.90 (s, 3H), 3.48 (s, 3H), 3.48 (s, 3H), 2.11-1.79 (m, 9H), 1.13-1.06 (m, 18H).

MS (ESI) m/z: 839 (M+1)⁺.

General procedure B (starting from 41) to afford 40 (16 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.03-8.99 (m, 1H), 7.63-7.60 (m, 2H), 7.52 (dd, J=8.0, 1.6 Hz, 1H), 7.39-7.27 (m, 2H), 7.18-7.15 (m, 2H), 6.85 (d, J=9.2 Hz, 1H), 4.53-4.36 (m, 3H), 3.93 (s, 3H), 3.48 (s, 6H), 1.80 (d, J=7.1 Hz, 3H), 1.74 (d, J=7.3 Hz, 3H), 1.71-1.68 (m, 3H).

MS (ESI) m/z: 713 (M+1)⁺.

General procedure E (starting from 156, 2 days) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 41 (58 mg, 92%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.3 Hz, 1H), 7.44-7.32 (m, 2H), 7.25-7.18 (m, 4H), 7.07 (d, J=7.5 Hz, 1H), 6.79 (d, J=7.5 Hz, 1H), 5.11-5.09 (m, 3H), 4.64-4.60 (m, 3H), 3.81 (s, 3H), 3.44 (s, 6H), 1.63-1.55 (m, 9H), 1.49 (s, 9H), 1.47 (s, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 171.5, 171.3, 171.1, 155.0, 154.8, 152.2, 150.8, 147.6, 145.3, 140.6, 140.0, 139.4, 134.4, 133.3, 128.0, 127.9, 123.9, 123.7, 123.7, 123.7, 123.6, 123.0, 120.6, 115.7, 115.1, 112.7, 112.2, 112.0, 108.9, 106.3, 106.1, 80.0 (3C), 56.2 (2C), 55.8, 55.7, 55.7, 55.5, 28.3 (9C), 18.6 (3C).

MS (ESI) m/z: 1035 (M+23)⁺, 1013 (M+1)⁺.

Rf: 0.43 (hexane:EtOAc, 50:50).

General procedure B (starting from 156) to afford 42 (17 mg, quant.)

¹H NMR (300 MHz, CD₃OD) δ 7.46-7.44 (m, 2H), 7.28-7.27 (m, 1H), 7.19 (s, 1H), 7.16 (s, 1H), 6.87 (d, J=8.8 Hz, 1H), 6.78 (d, J=10.0 Hz, 1H), 4.75 (t, J=6.2 Hz, 2H), 4.77-4.37 (m, 3H), 3.87 (s, 3H), 3.45 (s, 3H), 3.38 (s, 3H), 3.16 (t, J=6.2 Hz, 2H), 1.77 (d, J=6.9 Hz, 3H), 1.71-1.67 (m, 6H).

MS (ESI) m/z: 715 (M+1)⁺.

General procedure B (starting from 160) to afford 43 (13.0 mg, quant.)

¹H NMR (300 MHz, CD₃OD) δ 9.18 (d, J=7.6 Hz, 1H), 7.56-7.52 (m, 2H), 7.40-7.22 (m, 4H), 6.90 (d, J=10.7 Hz, 1H), 4.60-4.59 (m, 1H), 4.37-4.35 (m, 1H), 4.26-4.24 (m, 1H), 3.90 (br s, 6H), 3.54 (br s, 3H), 3.47 (s, 3H); 2.62-2.47 (s, 3H), 1.32-1.19 (s, 18H).

MS (ESI) m/z: 827 (M+1)⁺.

General procedure B (starting from 158) to afford 44 (12.3 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.20 (d, J=8.1 Hz, 1H); 7.55-7.47 (m, 2H), 7.27-7.18 (m, 19H), 6.87 (d, J=9.3 Hz, 1H), 5.03 (s, 6H), 4.65 (br t, 1H), 4.49 (br t, 1H), 4.36 (br t, 1H), 3.88 (br s, 3H), 3.85 (br s, 3H), 3.51 (br s, 3H), 3.46 (br s, 3H), 3.22-3.18 (m, 6H), 2.40-2.00 (m, 6H), 1.65-1.50 (m, 12H).

MS (ESI) m/z: 1339 (M+23)⁺, 1316 (M+1)⁺.

General procedure B (starting from 159) to afford 45 (27.3 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 9.20 (d, J=7.6 Hz, 1H), 7.56-7.51 (m, 2H), 7.40-7.22 (m, 4H), 6.89 (d, J=8.3 Hz, 1H), 4.64 (t, J=6.0 Hz, 1H), 4.44 (t, J=6.3 Hz, 1H), 4.34 (t, J=7.2 Hz, 1H), 3.89 (br s, 6H), 3.54 (br s, 3H), 3.48 (br s, 3H), 2.19-1.79 (m, 9H), 1.15-1.07 (m, 18H).

MS (ESI) m/z: 869 (M+1)⁺.

General procedure D (starting from 1 and Boc-Ala-OH) and chromatography on silica gel (hexane:EtOAc, 50:50) to afford 46 (80 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 7.28-7.20 (m, 1H), 7.20-7.05 (m, 3H), 6.67 (d, J=3.7 Hz, 1H), 6.65 (d, J=4.0 Hz, 1H), 5.15-5.05 (m, 2H), 4.70-4.50 (m, 3H), 3.80 (s, 3H), 3.78 (s, 3H), 3.40 (s, 3H), 3.38 (s, 3H), 3.01 (br t, 2H), 1.70-1.50 (m, 9H), 1.48 (s, 9H), 1.47 (s, 9H), 1.46 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.8, 171.4, 155.0, 152.1, 151.8, 147.5, 144.9, 144.8, 141.3, 141.0, 139.8, 138.7, 134.8, 134.7, 134.3, 127.1, 127.0, 123.7, 123.2, 122.7, 119.7, 119.4, 116.2, 115.7, 114.9, 114.7, 111.8, 107.6, 105.5, 80.2, 80.1 (2C), 60.8, 56.2, 55.7, 55.5, 49.5, 49.3 (2C), 41.9, 28.3 (9C), 22.1, 18.7, 18.6, 18.3.

MS (ESI) m/z: 1067 (M+23)⁺, 1045 (M+1)⁺.

Rf: 0.70 (hexane:EtOAc, 1:2).

General procedure D (starting from 95 and (+)-biotine) and chromatography on silica gel (CH₂Cl₂:MeOH, 10:1) to afford 47 (5 mg, 10%).

¹H NMR (300 MHz, CDCl₃) δ 7.30-7.10 (m, 4H), 6.80-6.60 (m, 3H), 6.40 (br s, 1H), 6.30 (br s, 1H), 6.20 (br s, 1H), 6.10 (br s, 1H), 4.90-4.70 (m, 2H), 4.60-4.45 (m, 2H), 4.40-4.25 (m, 2H), 3.90 (s, 3H), 3.80 (s, 3H), 3.45 (s, 3H), 3.39 (s, 3H), 3.20-3.10 (m, 4H), 3.00-2.50 (m, 8H), 2.00-1.50 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 171.6, 171.3, 163.8, 155.2, 152.1, 149.1, 147.6, 144.9, 139.8, 139.0, 135.7, 134.2, 127.4, 126.5, 123.8, 123.5, 123.3, 119.7, 116.0, 114.8, 114.4, 111.8, 111.0, 108.5, 105.4, 62.3, 62.2, 61.4, 60.3, 59.9, 56.2, 56.1, 55.9, 55.8, 55.5, 55.3, 55.2, 42.5, 40.6, 40.5, 33.8, 33.1, 29.7, 28.6, 28.2, 28.1, 27.8, 27.4, 25.1, 24.7, 21.0, 14.2.

MS (ESI) m/z: 990 (M+23)⁺, 968 (M+1)⁺.

Rf: 0.20 (CH₂Cl₂:MeOH, 10:1).

General procedure A (starting from 49) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 10:1) to afford 48 (9.9 mg, 84%).

¹H NMR (300 MHz, CDCl₃) δ 9.60 (br s, 1H), 7.15-7.07 (m, 2H), 7.00 (br s, 1H), 6.82 (s, 1H), 6.72 (s, 1H), 6.40 (s, 1H), 6.02 (br s, 1H), 5.80 (br s, 2H), 5.16-5.08 (m, 1H), 4.85-4.78 (m, 1H), 3.88 (s, 3H), 3.87 (s, 3H), 3.47 (s, 3H), 3.36 (s, 3H), 3.18-3.10 (m, 2H).

MS (ESI) m/z: 531 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂:MeOH, 10:1).

General procedure G (starting from 6,7-dimethoxy-5-isopropoxy-3,4-dihydroisoquinoline and 2-Bromo-N-[5-isopropoxy-2-(4-isopropoxy-3-methoxy-phenylethynyl)-4-methoxy-phenyl]-acetamide) and purification by chromatography on silica gel (EtOAc:hexane, 4:1) to afford 49 (55.7 mg, 21%).

¹H NMR (300 MHz, CDCl₃) δ 7.10-7.08 (m, 3H), 7.00 (s, 1H), 6.77 (s,1H), 6.63 (s, 1H), 4.99 (br t, 2H), 4.63-4.53 (m, 3H), 3.83 (s, 6H), 3.41 (s, 3H), 3.35 (s, 3H), 3.16 (br t, 2H), 1.42 (d, J=5.4 Hz, 6H), 1.40 (d, 5.4 Hz, 6H), 1.32 (d, 6.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 156.7, 151.7, 151.3, 148.5, 146.8, 145.9, 142.0, 133.3, 129.8, 129.7, 127.3, 123.7, 123.6, 121.0, 118.9, 117.0, 115.4, 114.9, 111.2, 105.3, 104.9, 102.6, 75.7, 71.8, 71.6, 60.5, 56.2, 55.3, 55.1, 42.3, 23.0, 22.7 (2C), 22.0 (2C), 21.9 (2C).

MS (ESI) m/z: 657 (M+1)⁺.

Rf: 0.34 (EtOAc:hexane, 4:1).

General procedure G (starting from 6-isopropoxy-7-methoxy-3,4-dihydroisoquinoline and 2-Bromo-N-[5-isopropoxy-2-(4-isopropoxy-3-methoxy-phenylethynyl)-4-methoxy-phenyl]-acetamide) and purification by chromatography on silica gel (EtOAc) to provide 50 (51.7 mg, 18%).

¹H NMR (300 MHz, CDCl₃) δ 7.09-7.08 (m, 3H), 6.95 (br s, 1H), 6.80-6.76 (m, 3H), 5.03-5.00 (m, 2H), 4.62-4.52 (m, 3H), 3.81 (s, 3H), 3.41 (s, 3H), 3.34 (s, 3H), 3.11 (t, J=6.3 Hz, 2H), 1.42-1.34 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 156.1, 151.1, 148.4, 146.5, 145.8, 134.0, 129.6, 129.3, 127.4, 126.3, 123.5, 120.7, 118.4, 116.9, 114.7, 114.4, 111.1, 109.0, 105.0, 102.3, 71.8, 71.4, 71.3, 55.9, 55.0, 54.9, 42.3, 28.7, 21.7 (6C).

MS (ESI) m/z: 627 (M+1)⁺.

Rf: 0.42 (EtOAc).

General procedure E (starting from 52) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 51 (7 mg, 64%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.3 Hz, 1H), 7.63 (d, J=1.2 Hz, 1H), 7.53 (dd, J=8.0, 1.7 Hz, 1H), 7.37 (d, J=1.5 Hz, 1H), 7.23-7.15 (m, 3H), 7.08 (d, J=7.3 Hz, 1H), 6.76 (s, 1H), 4.38-4.22 (m, 12H), 3.88 (s, 3H), 3.48 (s, 3H), 3.47 (s, 3H), 1.45 (t, J=7.0 Hz, 6H), 1.38 (t, J=7.0 Hz, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.1, 152.0 (d, J_(C-P)=4.5 Hz), 150.6 (d, J_(C-P)=5.5 Hz), 147.4 (d, J_(C-P)=5.0 Hz), 145.5, 140.9 (d, J_(C-P)=7.1 Hz), 140.2 (d, J_(C-P)=7.1 Hz), 139.9 (d, J_(C-P)=7.1 Hz), 133.5, 133.3, 128.2, 123.9, 123.7, 123.3, 122.8 (d, J_(C-P)=3.0 Hz), 122.6, 118.8, 115.4, 114.6, 112.8, 111.9, 110.8, 108.9, 106.5, 106.3, 64.9, 64.8 (2C), 64.7 (2C), 64.6, 56.3, 55.8, 55.6, 16.2, 16.1 (3C), 16.0.

MS (ESI) m/z: 908 (M+1)⁺.

Rf: 0.29 (CH₂Cl₂:MeOH, 20:1).

To a suspension of 109 (15 mg, 0.030 mmol) in anhydrous CH₂Cl₂ under Argon atmosphere, Et₃N (17 μL, 0.120 mmol) and diethyl chlorophosphate (18 μL, 0.120 mmol) were added and the mixture was stirred at 23° C. After 4.5 h, two more equivalents of Et₃N (9 μL, 0.060 mmol) and diethyl chlorophosphate (9 μL, 0.060 mmol) were added and the mixture stirred at 23° C. overnight. The mixture was concentrated under reduced pressure and the residue purified by chromatography on silica gel (CH₂Cl₂:MeOH, from 30:1 to 15:1) to give 52 as a white solid (20 mg, 74%).

¹H NMR (300 MHz, CDCl₃) δ 7.45 (dd, J=8.1, 1.5 Hz, 1H), 7.29 (d, J=1.5 Hz, 1H), 7.18 (s, 1H), 7.10 (dd, J=8.1, 1.6 Hz, 1H), 7.06 (s, 1H), 6.71 (s, 1H), 6.64 (s, 1H), 4.94-4.86 (m, 1H), 4.72-4.63 (m, 1H), 4.34-4.18 (m, 12H), 3.84 (s, 3H), 3.44 (s, 3H), 3.36 (s, 3H), 3.09 (br t, 2H), 1.44-1.31 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 155.1, 151.7 (d, J_(C-P)=4.5 Hz), 149.3 (d, J_(C-P)=5.5 Hz), 147.4 (d, J_(C-P)=5.0 Hz), 144.9, 139.9 (d, J_(C-P)=7.1 Hz), 139.6 (d, J_(C-P)=6.5 Hz), 139.1 (d, J_(C-P)=7.6 Hz), 135.0, 133.0, 127.0, 126.2, 124.4, 123.2, 122.6, 122.6, 121.1, 115.5, 114.9, 114.8, 110.5, 109.8, 105.6, 64.7, 64.7, 64.7 (3C), 64.6, 56.2, 55.8, 55.5, 42.4, 28.1, 16.2, 16.1 (3C), 16.0, 16.0.

MS (ESI) m/z: 910 (M+1)⁺.

Rf: 0.23 (CH₂Cl₂:MeOH, 30:1).

General procedure A (starting from 54) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, from 20:1 to 10:1) to afford 53 (70 mg, 51%).

¹H NMR (300 MHz, DMSO-d₆) δ 9.30 (d, J=7.6 Hz, 1H), 8.20 (d, J=7.7 Hz, 1H), 8.08 (d, J=8.0 Hz, 1H), 7.70-7.50 (m, 2H), 7.20-7.10 (m, 2H), 7.00 (d, J=7.8 Hz, 1H), 6.88 (s, 1H), 6.57 (s, 1H), 5.69 (s, 1H), 3.76 (s, 3H), 3.40 (s, 3H).

MS (ESI) m/z: 499 (M+1)⁺.

Rf: 0.61 (CH₂Cl₂:MeOH, 10:1).

General procedure H (starting from 5-nitroisoquinoline) and purification by chromatography on silica gel (hexane:CH₂Cl₂:Et₂O, from 5:5:1 to 5:5:2) to afford 54 (190 mg, 33%).

¹H NMR (300 MHz, CD₃OD) δ 9.41 (d, J=7.8 Hz, 1H), 8.12 (dd, J=7.8, 1.1 Hz, 1H), 8.06 (dt, J=8.0, 0.9 Hz, 1H), 7.78 (dd, J=7.8, 0.7 Hz, 1H), 7.36 (t, J=8.2 Hz, 1H), 7.19 (d, J=8.2 Hz, 1H), 7.11 (dd, J=8.2, 1.8 Hz, 1H), 7.05 (d, J=1.8 Hz, 1H), 6.96 (s, 1H), 6.63 (s, 1H), 4.71 (hp, J=6.0 Hz, 1H), 4.58 (hp, J=6.0 Hz, 1H), 3.84 (s, 3H), 3.44 (s, 3H), 1.51 (d, J=6.0 Hz, 3H), 1.44 (d, J=6.0 Hz, 3H), 1.40 (d, J=7.8 Hz, 6H).

MS (ESI) m/z: 583 (M+1)⁺.

Rf: 0.50 (hexane:CH₂Cl₂:Et₂O, 5:5:2).

General procedure E (starting from 28, reaction time 15 h) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 55 (17 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.5 Hz, 1H), 7.38 (s, 1H), 7.30-7.13 (m, 5H), 7.05 (d, J=7.3 Hz, 1H), 6.81 (s, 1H), 3.82 (s, 3H), 3.45 (s, 6H), 2.65-2.55 (m, 6H), 1.82-1.73 (m, 6H), 1.42-1.25 (m, 24H), 0.91-0.85 (m, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.7, 171.6, 171.6, 155.1, 152.4, 151.1, 147.8, 145.5, 141.0, 140.4, 139.9, 134.1, 133.6, 128.3, 124.1, 123.8, 123.6, 123.6, 123.1, 120.7, 115.6, 115.0, 112.8, 112.3, 112.2, 109.0, 106.4, 106.1, 56.2, 55.8, 55.6, 34.0 (3C), 31.7 (3C), 29.7, 29.0 (2C), 28.9 (3C), 25.0, 25.0, 24.9, 22.6 (3C), 14.1 (3C).

MS (ESI) m/z: 878 (M+1)⁺.

Rf: 0.31 (CH₂Cl₂:MeOH, 100:1).

A suspension of 86 (0.2248 g, 0.288 mmol), Pd(OAc)₂ (3.8 mg, 0.017 mmol), BINAP (16.2 mg, 0.026 mmol), and Cs₂CO₃ (0.263 g, 0.807 mmol) in anhydrous toluene (5 mL) was stirred at 23° C. under Argon atmosphere for 5 min. Then benzophenone imine (116 mL, 0.692 mmol) was added and the mixture was heated at 110° C. for 3 d. The reaction was cooled to 23° C., CH₂Cl₂ was added (20 mL), and washed with H₂O (20 mL).

The combined organic layers were dried over anhydrous Na₂SO₄, filtered and evaporated to dryness. The residue was purified by chromatography on silica gel (hexane:EtOAc, 50:50) to give LL-MA-triflate-NPh₂ (56.2 mg, 24%) and 56 (0.102 g, 42%) as a yellow solids.

¹H NMR (300 MHz, CDCl₃) δ 7.76-7.70 (m, 4H), 7.48-7.37 (m, 7H), 7.32-7.20 (m, 7H), 7.14-7.10 (m, 2H), 6.98 (dd, J=7.8, 1.5 Hz, 1H), 6.91 (d, J=1.5 Hz, 1H), 6.77 (d, J=7.8 Hz, 1H), 6.74 (s, 1H), 6.71 (s, 1H), 6.60 (s, 1H), 6.59 (s, 1H), 4.89-4.81 (m, 1H), 4.68-4.59 (m, 1H), 3.90 (s, 3H), 3.68 (s, 3H), 3.35 (s, 3H), 3.27 (s, 3H), 3.05 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 170.2, 169.2, 155.4, 150.1, 148.8, 147.4, 146.3, 145.4, 140.9, 140.7, 139.3, 138.9, 136.8, 136.3, 135.6, 130.8, 130.7, 130.6, 129.4, 129.3, 128.7, 128.6, 128.5, 128.4, 128.0, 127.7, 127.6, 126.6, 123.3, 120.9, 119.9, 115.1, 114.0, 113.9, 113.4, 110.8, 109.0, 108.9, 104.6, 55.8, 55.6, 55.5, 55.4, 42.2, 28.6.

MS (ESI) m/z: 842 (M+1)⁺.

Rf: 0.33 (hexane:EtOAc, 50:50).

HCl 1.5 N (1.5 mL) was added to a solution of 56 (91.0 mg, 0.108 mmol) in THF (20 mL) at 23° C. The solution turned from yellow to colorless in 10 min. The solvent was evaporated to dryness and H₂O was added (20 mL).

The suspension was basified with aqueous ammonia 32% (0.5 mL) and extracted with CH₂Cl₂ (3×20 mL), dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated to give a residue which was purified by chromatography on silica gel (hexane:EtOAc, 1:4) to give 57 as a white solid (55 mg, quant).

¹H NMR (300 MHz, CDCl₃) δ 6.95-6.85 (m, 3H), 6.78 (s, 1H), 6.72 (s, 1H), 6.69 (s, 1H), 6.64 (s, 1H), 4.84-4.78 (m, 1H), 4.71-4.65 (m, 1H), 3.98 (br s, 4H), 3.86 (s, 3H), 3.79 (s, 3H), 3.45 (s, 3H), 3.38 (s, 3H), 3.08 (br t, J=7.3 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 155.8, 148.6, 147.7, 147.2, 146.6, 143.7, 136.4, 135.8, 135.7, 128.9, 126.4, 124.9, 123.7, 120.3, 115.0, 113.1, 112.9, 110.8, 108.7, 108.3, 103.9, 102.1, 55.8, 55.6, 55.1 (2C), 42.2, 29.2, 28.6.

MS (ESI) m/z: 514 (M+1)⁺.

Rf: 0.32 (hexane:EtOAc, 1:4).

General procedure E (starting from 84, reaction time 20 h) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to give 58 (30.7 mg, 96%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.3 Hz, 1H), 7.39-7.25 (m, 11H), 7.10-7.05 (m, 6H), 6.82 (d, J=7.3 Hz, 1H), 5.03 (m, 2H), 4.91 (m, 2H), 3.99 (s, 3H), 3.84 (s, 3H), 3.49 (s, 3H), 3.44 (s, 3H), 3.37-3.23 (m, 4H), 1.45 (s, 9H), 1.42 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 170.0, 154.9, 152.1, 150.3, 149.5, 147.5, 145.4, 139.8, 139.2, 135.7 (2C), 134.8, 134.2 (2C), 129.5, 128.6, 128.1 (2C), 124.7, 123.8, 123.0, 119.0, 116.0, 115.4, 113.0, 112.0, 110.8, 108.4, 107.4, 106.2, 105.1, 80.1 (2C), 56.1, 55.9, 55.6 (2C), 54.4 (2C), 38.1 (2C), 28.2 (6C).

MS (ESI) m/z: 1008 (M+1)⁺.

Rf: 0.60 (CH₂Cl₂:MeOH, 60:1).

General procedure B (starting from 60) to afford 59 (15 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.50-7.35 (m, 2H), 7.30-7.20 (m, 2H), 6.85-6.75 (m, 2H), 4.80-4.65 (m, 2H), 4.60 (br s, 1H), 4.53 (t, J=3.8 Hz, 1H), 4.42 (br t, 1H), 4.30-4.05 (m, 6H), 3.87 (s, 3H), 3.79 (s, 3H), 3.43 (s, 3H), 3.42 (s, 3H), 3.06 (br s, 2H).

MS (ESI) m/z: 793 (M+1)⁺.

General procedure J (starting from 68, overnight) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 10:1) to afford 60 (23 mg, 82%).

¹H NMR (300 MHz, CDCl₃) δ 7.40-7.00 (m, 4H), 6.75-6.60 (m, 2H), 5.70-5.40 (m, 3H), 4.90-4.50 (m, 4H), 4.40-4.20 (m, 3H), 4.10-3.80 (m, 10H), 3.43 (s, 3H), 3.39 (s, 3H), 3.00 (br s, 2H), 2.80-2.50 (m, 3H), 1.49 (s, 9H), 1.46 (s, 9H), 1.45 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 169.4, 169.1, 155.6, 154.7, 151.5, 151.4, 147.4, 146.7, 146.4, 145.7, 145.1, 143.5, 141.1, 140.4, 139.6, 139.4, 138.4, 135.0, 134.7, 134.6, 126.9, 126.7, 126.5, 123.9, 123.8, 123.7, 122.9, 119.5, 116.3, 115.6, 114.9, 114.8, 114.3, 113.1, 112.1, 109.7, 107.8, 105.5, 103.5, 80.5, 64.0, 63.7, 61.0, 56.5, 56.3, 56.1, 55.8, 55.6, 55.5, 55.3, 41.9, 28.3, 22.1.

MS (ESI) m/z: 1115 (M+23)⁺, 1093 (M+1)⁺.

Rf: 0.45 (CH₂Cl₂:MeOH, 10:1).

General procedure E (starting from 108, reaction time 24 h) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 40:1) to afford 61 (12 mg, 60%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.3 Hz, 1H), 7.39 (s, 1H), 7.30 (d, J=7.7 Hz, 1H), 7.25-7.22 (m, 3H), 7.14 (s, 1H), 7.05 (d, J=7.5 Hz, 1H), 6.81 (s, 1H), 3.84 (s, 3H), 3.45 (s, 6H), 2.37 (s, 3H), 2.34 (s, 3H), 2.32 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 168.9, 168.7, 168.7, 155.0, 152.4, 151.0, 147.8, 145.4, 140.9, 140.3, 139.7, 134.2, 133.5, 128.2, 124.1, 123.8, 123.7, 123.6, 123.1, 120.7, 115.7, 115.0, 112.8, 112.3, 112.2, 109.1, 106.4, 106.1, 56.2, 55.7, 55.6, 20.6 (3C).

MS (ESI) m/z: 626 (M+1)⁺.

Rf: 0.40 (CH₂Cl₂:MeOH, 100:1).

General procedure D (starting from 109 and coumarin 3-carboxylic acid) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 40:1) to afford 62 (41 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 8.80 (s, 1H), 8.79 (s, 1H), 8.75 (s, 1H), 7.72-7.65 (m, 6H), 7.42-7.34 (m, 7H), 7.26-7.21 (m, 3H), 7.23 (s, 1H), 6.85 (s, 1H), 6.81 (s, 1H), 4.93-4.86 (m, 1H), 4.78-4.69 (m, 1H), 3.84 (s, 3H), 3.50 (s, 3H), 3.44 (s, 3H), 3.15 (br t, 2H).

MS (ESI) m/z: 1040 (M+23)⁺.

Rf: 0.24 (CH₂Cl₂:MeOH, 50:1).

General procedure B (starting from 21) to afford 63 (31.9 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.44-7.38 (m, 2H), 7.24-7.20 (m, 2H), 6.94 (br s, 1H), 6.82-6.78 (m, 1H), 6.71-6.68 (m, 1H), 4.72 (m, 1H), 4.49-4.38 (m, 2H), 3.44 (s, 3H), 3.35 (s, 6H), 3.30 (s, 3H), 3.13 (br t, 2H), 1.77 (br d, 3H), 1.70 (br d, 3H).

MS (ESI) m/z: 658 (M+1)⁺.

General procedure E (starting from 22, reaction time 77 h) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 80:1) to afford 64 (17 mg, 68%).

¹H NMR (300 MHz, CDCl₃) δ 9.16 (d, J=7.5 Hz, 1H), 7.66 (s, 1H), 7.60 (d, J=8.6 Hz, 1H), 7.33-7.30 (m, 3H), 7.18 (s, 1H), 7.06 (d, J=7.7 Hz, 1H), 6.76 (s, 1H), 3.96 (s, 3H), 3.49 (s, 6H), 3.37 (s, 3H), 3.24 (s, 3H), 3.21 (s, 3H).

MS (APCI) m/z: 734 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 80:1).

General procedure D (starting from 95 and (L)-N-Boc-valine) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 65 (83.6 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 7.25-7.09 (m, 4H), 6.76 (br s, 1H), 6.71-6.66 (m, 2H), 5.06 (br d, J=9.3 Hz, 1H), 4.91-4.71 (m, 2H), 4.53-4.50 (m, 2H), 3.89 (s, 3H), 3.77 (s, 3H), 3.40 (s, 6H), 3.13 (t, J=7.3 Hz, 2H), 2.41-2.37 (m, 2H), 1.47 (d, J=6.1 Hz, 18H), 1.12-0.99 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 170.3, 155.6 (2C), 154.9, 151.9, 149.1, 147.6, 147.4, 144.8, 139.5, 138.4, 135.7, 134.4, 127.1, 127.0, 126.4, 123.7, 123.3, 119.5, 116.2, 114.8, 114.6, 114.4, 111.7, 110.9, 108.5, 105.4, 79.9 (2C), 58.5 (2C), 55.9, 55.8, 55.5, 55.4, 42.4, 31.2, 31.1, 28.5, 28.2 (6C), 19.1, 18.9, 17.1, 17.0.

MS (ESI) m/z: 936 (M+23)⁺, 914 (M+1)⁺.

Rf: 0.22 (CH₂Cl₂:MeOH, 60:1).

General procedure B (starting from 68) to afford 66 (18 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.50-7.20 (m, 17H), 7.16-7.12 (m, 2H), 6.80-6.75 (m, 2H), 4.80-4.60 (m, 11H), 4.35-3.95 (m, 6H), 3.78 (s, 3H), 3.76 (s, 3H), 3.39 (d, J=2.1 Hz, 3H), 3.36 (d, J=2.1 Hz, 3H), 2.91 (br s, 2H).

MS (ESI) m/z: 1063 (M)⁺.

General procedure E (starting from 114, reaction time 70 h) and purification by chromatography on silica gel (hexane:EtOAc, 1:1) to afford 67 (52 mg, 81%).

¹H NMR (300 MHz, CDCl₃) δ 9.06 (d, J=7.5 Hz, 1H), 8.10-8.00 (m, 3H), 7.80-7.65 (m, 6H), 7.60-7.50 (m, 3H), 7.50-7.10 (m, 21H), 6.74 (s, 1H), 5.80-5.50 (m, 3H), 5.20-5.00 (m, 3H), 3.91 (s, 3H), 3.81 (s, 3H), 3.80-3.60 (m, 6H), 3.50-3.40 (m, 6H), 1.49 (s, 18H), 1.45 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 168.8, 168.4, 168.3, 168.2, 155.1, 154.9, 154.6, 153.1, 153.0, 152.0, 147.3, 145.3, 141.5, 140.2, 140.1, 140.0, 139.6, 139.0, 138.5, 138.1, 138.0, 136.7, 134.8, 132.9, 132.7, 132.5, 128.0, 127.9, 127.8, 127.7, 127.5, 127.3, 127.2, 127.1, 127.0, 126.9, 126.8, 126.7, 125.9, 125.3, 125.2, 123.8, 123.6, 123.5, 120.8, 119.9, 119.8, 119.7, 119.4, 119.2, 119.1, 117.9, 116.0, 115.1, 112.0, 111.9, 109.0, 106.6, 106.1, 104.4, 80.9, 80.8, 80.7, 61.0, 56.2, 55.8, 55.7, 55.6, 54.3, 54.1, 38.8, 29.7, 28.3, 28.2, 27.3.

MS (ESI) m/z: 1689 (M+23)⁺.

Rf: 0.19 (hexane:EtOAc, 2:1).

General procedure D (starting from 1 and (L)-N-Boc-Ser(Bzl)) and purification by chromatography on silica gel (hexane:EtOAc, 50:50) to afford 68 (153 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 7.40-7.25 (m, 15H), 7.20-7.05 (m, 4H), 6.70-6.65 (m, 2H), 5.60-5.45 (m, 3H), 4.80-4.50 (m, 11H), 4.20-4.00 (m, 3H), 3.90-3.80 (m, 3H), 3.74 (s, 6H), 3.36 (t, J=4.7 Hz, 6H), 2.89 (br s, 2H), 1.48 (s, 18H), 1.46 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 169.0, 168.8, 155.4, 155.3, 154.9, 152.1, 151.8, 147.5, 144.8, 141.3, 141.1, 139.7, 138.6, 137.4, 137.0, 134.7, 134.3, 128.6, 128.5, 128.4, 128.2, 127.8, 127.6, 127.5, 126.9, 123.8, 123.1, 122.8, 119.4, 116.1, 115.6, 114.7, 111.8, 107.6, 105.5, 80.2, 80.1, 80.0, 73.7, 73.4, 70.0, 69.9, 60.8, 60.3, 56.1, 55.7, 55.5, 54.2, 54.1, 54.0, 41.8, 28.2, 22.0.

MS (ESI) m/z: 1385 (M+23)⁺.

Rf: 0.59 (hexane:EtOAc, 50:50).

General procedure G (starting from 6,7-dimethoxy-3,4-dihydroisoquinoline and 2-bromo-N-[5-isopropoxy-2-(4-isopropoxy-3-methoxy-phenylethynyl)-4-methoxy-phenyl]-acetamide) and purification by chromatography on silica gel (EtOAc) to afford 69 (4.2 mg, 9%).

¹H NMR (300 MHz, CDCl₃) δ 9.71 (br s, 1H), 7.10 (br s, 2H), 7.07 (s, 1H), 6.81 (s, 1H), 6.78 (s, 1H), 6.76 (m, 2H), 5.03-4.94 (m, 2H), 4.61-4.55 (m, 2H), 3.89 (s, 3H), 3.82 (s, 3H), 3.41 (s, 3H), 3.37 (s, 3H), 3.12 (t, J=6.8 Hz, 2H), 1.40 (d, J=5.9 Hz, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 156.6, 151.2, 148.3, 147.3, 146.6, 145.8, 133.6, 129.7, 127.3, 126.3, 123.5, 120.7, 118.6, 116.9, 114.8, 114.5, 111.1, 110.9, 108.5, 105.1, 102.5, 71.7, 71.4, 56.0, 55.8, 55.1, 55.0, 42.3, 29.0, 21.9 (2C), 20.9 (2C).

MS (ESI) m/z: 599 (M+1)⁺.

Rf: 0.21 (EtOAc).

General procedure A (starting from 69) and purification by chromatography on silica gel (EtOAc) to afford 70 (2.4 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 9.26 (br s, 1H), 7.12-7.07 (m, 2H), 7.00 (s, 1H), 6.79-6.73 (m, 4H), 5.75 (s, 2H), 5.15-5.11 (m, 1H), 4.81 (m, 1H), 3.89 (s, 3H), 3.86 (s, 3H), 3.49 (s, 3H), 3.39 (s, 3H), 3.13-3.11 (m, 2H).

MS (ESI) m/z: 515 (M+1)⁺.

Rf: 0.41 (CH₂Cl₂:MeOH, 10:1).

General procedure B (starting from 74) to afford 71 (14.0 mg, quant.)

¹H NMR (300 MHz, CD₃OD) δ 7.45-7.22 (m, 19H), 6.80-6.74 (m, 2H), 5.08 (m, 1H), 5.03 (s, 6H), 4.78 (m, 1H), 4.51 (br t, 1H), 4.42 (br t, 1H), 4.35 (br t, 1H), 3.79 (s, 6H), 3.42 (s, 3H), 3.41 (s, 3H), 3.21 (br s, 6H), 3.05 (m, 2H), 2.15 (m, 6H), 1.65 (m, 12H).

MS (ESI) m/z: 1340 (M+23)⁺, 1318 (M+1)⁺.

General procedure B (starting from 75) to afford 72 (14.1 mg, quant.)

¹H NMR (300 MHz, CD₃OD) δ 7.48-7.42 (m, 2H), 7.28-7.20 (m, 2H), 6.83-6.77 (m, 2H), 4.76 (m, 2H), 4.45 (dd, J=4.4, 2.0 Hz, 1H), 4.33 (dd, J=4.4, 2.0 Hz, 1H), 4.23 (dd, J=4.4, 2.0 Hz, 1H), 3.87 (s, 3H), 3.82 (s, 3H), 3.44 (s, 3H), 3.43 (s, 3H), 3.05 (m, 2H), 2.59-2.44 (m, 3H), 1.38-1.20 (m, 18H).

MS (ESI) m/z: 829 (M+1)⁺.

Rf: 0.28 (CH₂Cl₂:MeOH, 10:1).

General procedure B (starting from 76) to afford 73 (13.8 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.47-7.41 (m, 2H), 7.31-7.21 (m, 2H), 6.81-6.77 (m, 2H), 4.90-4.76 (m, 2H), 4.48 (t, J=7.3 Hz, 1H), 4.40 (t, J=7.1 Hz, 1H), 4.42 (t, J=6.6 Hz, 1H), 3.86 (s, 3H), 3.80 (s, 3H), 3.44 (s, 3H), 3.42 (s, 3H), 3.17-3.06 (m, 2H), 2.14-1.77 (m, 9H), 1.12-1.00 (m, 18H).

MS (ESI) m/z: 871 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂:MeOH, 10:1).

General procedure D (starting from 1 and (L)-N-Boc-Lysine-CBz) and purification by chromatography on silica gel (hexane:EtOAc, 2:3) to afford 74 (114.6 mg, 75%).

¹H NMR (300 MHz, CDCl₃) δ 7.34-7.31 (m, 15H), 7.15-7.07 (m, 4H), 6.67-6.63 (m, 2H), 5.08 (br s, 6H), 4.92 (m, 3H), 4.55 (m, 2H), 3.77 (s, 6H), 3.39 (s, 3H), 3.37 (s, 3H), 3.26-3.17 (m, 6H), 3.00 (br t, 2H), 2.04-1.76 (m, 6H), 1.58-1.29 (m, 38H).

¹³C NMR (75 MHz, CDCl₃) δ 171.0, 170.7 (2C), 156.5, 156.4, 155.5, 155.3 (2C), 154.8, 152.0, 151.7, 147.4 (2C), 144.8, 144.7, 141.2 (2C), 140.9 (2C), 139.6, 138.4 (2C), 136.5, 136.4, 134.7, 134.6, 134.2, 128.4, 128.0, 126.9, 126.8, 123.7, 123.2, 122.6, 119.4 (2C), 116.0, 115.6, 114.7(2C), 111.7, 110.8, 105.4 (2C).

MS (ESI) m/z: 1640 (M+23)⁺.

Rf: 0.30 (hexane:EtOAc, 2:3).

General procedure D (starting from 1 and (D)-N-Boc-Valine) and purification by chromatography on silica gel (hexane:EtOAc, 2:1) to afford 75 (96.5 mg, 91%).

¹H NMR (300 MHz, CDCl₃) δ 7.23 (s, 1H), 7.17-7.15 (m, 1H), 7.10-7.09 (m, 2H), 6.69-6.40 (m, 2H), 5.05 (br t, J=8.1 Hz, 2H), 4.53-4.51 (m, 3H), 3.78 (s, 6H), 3.40 (s, 3H), 3.38 (s, 3H), 3.01 (br t, 2H), 2.40-2.38 (m, 3H), 1.49 (br s, 27H), 1.15-0.99 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.5, 170.3, 155.7, 155.6 (2C), 154.9, 152.0, 151.8, 147.5, 144.8, 141.2, 141.0, 139.6, 138.5, 134.8, 134.3, 127.0, 126.9, 123.8, 123.2, 122.6, 119.4, 116.1, 115.6, 114.8, 114.7, 111.8, 107.6, 105.4, 80.1, 79.9 (2C), 58.9, 58.5 (2C), 56.0 (2C), 55.5 (2C), 41.8, 31.2, 31.1, 30.8, 28.2 (9C), 22.2, 19.2, 19.1, 18.9, 17.4, 17.1, 17.0.

MS (ESI) m/z: 1129 (M+1)⁺.

Rf: 0.23 (hexane:EtOAc, 2:1).

General procedure D (starting from 1 and (L)-N-Boc-Leucine) and purification by chromatography on silica gel (hexane:EtOAc, 2:1) to afford 76 (100.1 mg, 91%).

¹H NMR (300 MHz, CDCl₃) δ 7.24-7.08 (m, 4H), 6.68-6.64 (m, 2H), 4.92 (m, 2H), 4.57 (m, 3H), 3.79 (s, 3H), 3.77 (s, 3H), 3.40 (s, 3H), 3.39 (s, 3H), 3.02 (br t, 2H), 1.87-1.85 (m, 2H), 1.87-1.85 (m, 3H), 1.70-1.61 (m, 3H), 1.58 (br s, 27H), 1.05-0.98 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4 (2C), 155.5, 155.4, 155.3, 155.0, 152.1, 151.8, 147.5, 144.8, 141.4, 141.0, 139.8, 138.7, 134.8, 134.2, 127.1, 123.8, 123.2, 122.7, 119.5, 116.1, 115.7, 114.7 (2C), 111.8, 107.6, 105.4 (2C), 80.2, 80.0 (2C), 60.7, 56.1, 55.7, 55.5, 52.3 (3C), 41.9, 41.6, 41.5, 41.2, 29.6, 28.3 (9C), 24.7 (3C), 22.9 (2C), 22.8 (2C), 22.7, 21.8.

MS (ESI) m/z: 1193 (M+23)⁺, 1171 (M+1)⁺.

Rf: 0.29 (hexane:EtOAc, 2:1).

General procedure D (starting from 26 and (L)-N-Boc-Alanine) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 77 (12.4 mg, 62%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.6 Hz, 1H), 7.34-7.08 (m, 7H), 6.74 (d, J=8.1 Hz, 1H), 5.30-5.12 (m, 2H), 4.62-4.60 (m, 2H), 4.00 (s, 3H), 3.80 (s, 3H), 3.51 (s, 3H), 3.44 (s, 3H), 1.64-1.44 (m, 24H).

¹³C NMR (75 MHz, CDCl₃) δ 171.5, 155.0, 152.2, 150.3, 149.6, 147.5, 145.5, 140.0, 139.4, 134.8, 134.2 (2C), 128.3 (2C), 124.7, 123.8, 123.2. 119.0, 116.0, 115.3, 113.0, 112.0, 110.9, 108.4, 107.5, 106.2, 105.1, 80.1, 56.2, 56.0, 55.8, 55.7, 49.3, 28.3, 18.7.

MS (ESI) m/z: 878 (M+23)⁺, 856 (M+1)⁺.

Rf: 0.13 (CH₂Cl₂:MeOH, 60:1).

General procedure D (starting from 26 and (D)-N-Boc-Valine) and purification by chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 78 (15.4 mg, 86%).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.3 Hz, 1H), 7.32-7.22 (m, 3H), 7.14-7.07 (m, 4H), 6.68 (d, J=8.8 Hz, 1H), 5.09-5.06 (m, 2H), 4.57-4.52 (m, 2H), 3.99 (s, 3H), 3.80 (s, 3H), 3.51 (s, 3H), 3.43 (s, 3H), 2.46-2.38 (m, 2H), 1.49-1.45 (m, 18H), 1.27-1.00 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4, 155.7, 155.0, 152.2, 150.3, 149.6, 147.5, 145.5, 139.9, 139.3, 134.9, 130.3, 134.8, 134.2, 128.2, 124.7, 123.7, 123.8, 123.1, 119.0, 116.0, 115.3, 113.0, 112.1, 110.9, 108.4, 107.5, 106.2, 105.1, 80.0 (2C), 58.6, 58.5, 56.0, 55.7, 55.6, 55.5, 31.3, 31.2, 28.3 (6C), 19.2, 19.1, 17.2, 17.1.

MS (ESI) m/z: 934 (M+23)⁺, 912 (M+1)⁺.

Rf: 0.32 (CH₂Cl₂:MeOH, 60:1).

General procedure D (starting from 26 and 9H-fluorene-4-carboxilic acid) and purification by chromatography on silica gel (hexane:EtOAc, from 2:1 to 1:1) to afford 79 (8.6 mg, 41%).

¹H NMR (300 MHz, CDCl₃) δ 9.31 (d, J=7.3 Hz, 1H), 8.53-8.45 (m, 2H), 8.21 (d, J=7.8 Hz, 1H), 8.20 (d, J=8.0 Hz, 1H), 7.71 (t, J=7.1 Hz, 2H), 7.61-7.53 (m, 3H), 7.47-7.32 (m, 10H), 7.16-7.13 (m, 2H), 7.05 (s, 1H), 4.02 (s, 3H), 3.99 (s, 2H), 3.96 (s, 2H), 3.90 (s, 3H), 3.66 (s, 3H), 3.61 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 166.0, 165.9, 155.2, 152.6, 150.4, 149.6, 148.0, 145.7, 145.2, 145.1, 144.3, 144.2, 141.6, 141.4, 140.5, 140.0, 139.9 (2C), 134.8, 134.3, 129.6, 129.4, 129.1 (2C), 128.5, 127.8, 127.7, 126.9, 126.7, 126.1, 126.0, 125.4, 125.1, 125.0, 124.9, 124.8 (2C), 124.6, 124.2, 124.0, 123.2, 119.1, 116.1, 115.4, 113.0, 112.4, 111.2, 108.5, 107.5, 106.4, 105.2, 56.3, 56.0, 55.9, 55.9, 37.0 (2C).

MS (ESI) m/z: 898 (M+1)⁺.

Rf: 0.50 (hexane:EtOAc, 50:50).

General procedure D (starting from 26 and 2[(4-fluorophenyl)thio]acetic acid) and purification by chromatography on silica gel (hexane:EtOAc, 2:1) to afford 80 (20.4 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.3 Hz, 1H), 7.58-7.50 (m, 4H), 7.26-7.20 (m, 4H), 7.11-7.01 (m, 8H), 6.78 (s, 1H), 4.00 (s, 3H), 3.87 (s, 2H), 3.81 (s, 2H), 3.77 (s, 3H), 3.48 (s, 3H), 3.37 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 167.5, 164.2, 161.0, 154.9, 152.1, 150.3, 149.5, 147.5, 145.4, 139.9, 139.3, 134.9, 134.2, 133.9, 133.8, 133.8, 133.7, 129.3, 128.1, 125.0, 124.7, 123.8, 123.6, 123.1, 118.9, 116.4, 116.1, 115.4, 113.0, 111.9, 110.8, 108.4, 107.5, 106.2, 105.0, 56.2, 56.0, 55.6, 55.5, 37.5, 37.4.

MS (ESI) m/z: 872 (M+23)⁺, 850 (M+1)⁺.

Rf: 0.52 (hexane:EtOAc, 2:1).

General procedure K (starting from 95 and 9H-fluorene-4-carboxilic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 81 as a yellow solid (20.0 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 8.53-8.43 (m, 2H), 8.17 (m, 2H), 7.78-7.41 (m, 2H), 7.30-7.10 (m, 12H), 6.94 (s, 1H), 6.83 (s, 1H), 6.80 (s, 1H), 4.96-4.80 (m, 2H), 3.98 (s, 2H), 3.96 (s, 2H), 3.93 (s, 3H), 3.88 (s, 3H), 3.60 (s, 3H), 3.54 (s, 3H), 3.19 (t, J=6.1 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 165.9, 155.2, 152.5, 149.2, 147.9, 147.8, 145.2, 145.1, 144.3, 144.1, 141.5, 141.3, 140.3, 139.9, 139.1, 136.0, 134.5, 129.5, 129.3, 129.0, 127.7, 127.6, 127.4, 127.3, 127.2, 126.8, 126.7, 126.6, 126.5, 126.1, 126.0, 125.9, 125.4, 125.2, 125.1, 125.0, 124.7, 124.5, 124.0, 123.5, 121.5, 119.7, 116.3, 115.0, 114.5, 112.1, 111.0, 108.6, 105.7, 56.2, 55.9, 55.9, 55.6, 42.6, 37.0 (2 C), 28.6.

MS (ESI) m/z: 922 (M+23)⁺, 900 (M+1)⁺.

Rf: 0.44 (CH₂Cl₂:MeOH, 200:1).

General procedure K (starting from 95 and 2,3,4,5-tetrafluorobenzoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 82 as a yellow solid (20.7 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 7.81-7.76 (m, 2H), 7.35 (d, J_(H-F)=7.8 Hz, 1H), 7.25-7.20 (m, 3H), 6.79 (br s, 2H), 6.71 (s, 1H), 4.92-4.79 (m, 2H), 3.91 (s, 3H), 3.82 (s, 3H), 3.48 (s, 3H), 3.46 (s, 3H), 3.16 (t, J=7.3 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 159.7 (2C), 155.0, 152.0, 150.2 (m), 149.3, 148.2 (m), 147.7, 147.4, 146.7 (m), 145.8 (m), 145.0 (m), 144.9, 143.4 (m), 142.3 (m), 140.0 (m), 139.4, 138.2, 136.0, 135.0, 127.0, 126.6, 123.6, 123.5, 119.5, 116.7, 115.1, 114.7, 114.6, 113.6 (m), 111.8, 111.0, 108.5, 105.6, 56.3, 55.9, 55.8, 55.5, 42.5, 28.6.

MS (ESI) m/z: 889 (M+23)⁺, 867 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂:MeOH, 200:1).

General procedure K (starting from 95 and (L)-N-Boc-Tryptophane) and chromatography on silica gel (CH₂Cl₂:MeOH, 25:1) to afford 83 as a yellow solid (13.0 mg, 98%).

¹H NMR (300 MHz, CDCl₃) δ 8.25-8.23 (br d, 2H), 7.68-7.63 (m, 2H), 7.40-7.34 (m, 2H), 7.26-7.05 (m, 7H), 6.93 (s, 1H), 6.77 (s, 1H), 6.67 (br s, 2H), 4.95-4.74 (m, 4H), 3.90 (s, 3H), 3.75 (s, 3H), 3.57-3.50 (m, 4H), 3.82 (s, 3H), 3.36 (s, 3H), 3.13 (br t, J=6.6 Hz, 2H), 1.43 (br d, 18H).

MS (ESI) m/z: 1110 (M+23)⁺.

Rf: 0.11 (CH₂Cl₂:MeOH, 30:1).

General procedure K (starting from 95 and (L)-N-Boc-Phenylalanine) and chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 84 as a yellow solid (13.0 mg, 96%).

¹H NMR (300 MHz, CDCl₃) δ 7.36-7.20 (m, 9H), 7.17-7.04 (m, 5H), 6.77 (s, 1H), 6.79-6.66 (m, 2H), 5.02-4.74 (m, 4H), 3.90 (s, 3H), 3.80 (s, 3H), 3.40 (s, 3H), 3.39 (s, 3H), 3.34-3.12 (m, 6H), 1.44-138 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 169.9, 154.9, 151.9, 149.1, 147.6, 147.4, 144.7, 139.4, 138.3, 135.8, 134.4, 129.4, 129.2, 128.5, 127.1, 126.4, 126.3, 123.6, 123.3, 119.5, 116.3, 114.9, 114.6, 114.4, 111.7, 110.9, 108.4, 105.4, 79.9 (2C), 56.0, 55.8, 55.6, 55.4, 54.3 (2C), 42.4, 38.0 (2C), 28.4, 28.2 (6C).

MS (ESI) m/z: 1032 (M+23)⁺.

Rf: 0.76 (CH₂Cl₂:MeOH, 60:1).

General procedure K (starting from 95 and 4-pentynoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 60:1) to afford 85 as a yellow solid (9.0 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.26-7.21 (m, 2H), 7.15-7.10 (m, 2H), 6.76 (s, 1H), 6.71 (s, 1H), 6.68 (s, 1H), 4.89-4.30 (m, 2H), 3.89 (s, 3H), 3.42 (s, 3H), 3.42 (s, 3H), 3.40 (s, 3H), 3.23 (t, J=7.1 Hz, 2H), 2.89-2.80 (m, 4H), 2.69-2.59 (m, 4H), 2.05-2.03 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 169.7, 169.5, 155.1, 152.1, 149.1, 147.7, 147.6, 144.9, 139.8, 138.7, 135.9, 134.4, 130.9, 128.8, 127.2, 126.4, 123.8, 123.3, 119.6, 116.2, 114.8, 114.5, 111.9, 111.0, 108.5, 105.5, 82.1, 82.0, 69.3 (2C), 56.2, 55.9, 55.7, 55.4, 42.5, 33.1 (2C), 28.6.

MS (ESI) m/z: 698 (M+23)⁺, 676 (M+1)⁺.

Rf: 0.65 (CH₂Cl₂:MeOH, 60:1).

General procedure I (starting from 95) and chromatography on silica gel (CH₂Cl₂) to give 86 as a yellow solid (13.4 mg, 89%).

¹H NMR (300 MHz, CDCl₃) δ 7.45 (d, J=8.8 Hz, 1H), 7.27 (s, 1H), 7.24-7.20 (m, 2H), 6.79 (s, 1H), 6.66 (s, 1H), 6.54 (s, 1H), 4.93-4.86 (m, 1H), 4.77-4.70 (m, 1H), 3.92 (s, 3H), 3.90 (s, 3H), 3.47 (s, 3H), 3.36 (s, 3H), 3.15 (br t, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 154.4, 152.5, 149.6, 147.8 (2C), 144.4, 138.5, 137.0 (3C), 136.2, 129.7, 126.8, 126.1, 123.6, 123.5, 118.6 (q, J_(C-F)=136.7 Hz), 118.6 (q, J_(C-F)=132.2 Hz), 115.7, 114.9, 114.0, 111.9, 111.2, 108.3, 105.7, 56.6, 56.0, 55.8, 55.2, 42.6, 28.5.

MS (ESI) m/z: 780 (M+1)⁺.

Rf: 0.43 (CH₂Cl₂).

General procedure K (starting from 167 and 4-methylcinnamic acid) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 87 as a white solid (5.5 mg, 86%).

¹H NMR (300 MHz, CDCl₃) δ 7.85 (d, J=16.4 Hz, 1H), 7.79 (d, J=16.8 Hz, 1H), 7.75 (d, J=15.8 Hz, 1H), 7.52-7.47 (m, 4H), 7.43-7.40 (m, 2H), 7.28-7.10 (m, 10H), 6.91 (s, 1H), 6.87 (s, 1H), 6.77 (s, 1H), 6.60 (d, J=16.1 Hz, 1H), 6.58 (d, J=16.1 Hz, 1H), 6.57 (d, J=16.1 Hz, 1H), 4.96-4.90 (m, 1H), 4.82-4.78 (m, 1H), 3.87 (s, 3H), 3.81 (s, 3H), 3.50 (s, 3H), 3.21 (t, J=7.1 Hz, 2H), 2.39 (s, 9H).

MS (ESI) m/z: 956 (M+23)⁺, 934 (M+1)⁺.

Rf: 0.45 (hexane:EtOAc, 1:1).

General procedure K (starting from 167 and cyclohexilpropionic acid) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 88 as a yellow oil (4.5 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 7.19-7.15 (m, 1H), 7.08-7.03 (m, 3H), 6.86 (s, 1H), 6.72-6.69 (m, 2H), 4.93-4.87 (m, 1H), 4.76-4.71 (m, 1H), 3.84 (s, 3H), 3.78 (s, 3H), 3.43 (s, 3H), 3.17 (br t, 2H), 2.64-2.55 (m, 6H), 1.77-1.40 (m, 21H), 1.36-1.02 (m, 12H), 0.98-0.88 (m, 6H).

MS (ESI) m/z: 938 (M+23)⁺, 916 (M+1)⁺.

Rf: 0.63 (hexane:EtOAc, 1:1).

General procedure K (starting from 167 and coumarin-3-carboxylic acid), chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) and the yellow solid was triturated with MeOH to afford 89 as a bright yellow solid (9.2 mg, 86%).

¹H NMR (300 MHz, CDCl₃) δ 8.78 (s, 1H), 8.75 (s, 1H), 8.70 (s, 1H), 7.77-7.62 (m, 6H), 7.37-7.33 (m, 6H), 7.24-7.13 (m, 4H), 6.93 (s, 1H), 6.87 (s, 1H), 6.80 (s, 1H), 4.92 (m, 1H), 4.84 (m, 1H), 3.87 (s, 3H), 3.82 (s, 3H), 3.50 (s, 3H), 3.23 (br t, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 160.8, 160.4 (2C), 156.5 (2C), 155.5, 155.4, 155.1, 152.1, 151.2, 150.3 (2C), 147.7, 144.9, 139.7, 138.6, 138.2, 134.9 (4C), 134.7, 133.9, 133.3, 130.2, 129.8 (2C), 127.1, 125.0, 124.9, 124.1, 123.2, 120.3, 120.1, 117.9, 117.8, 117.1, 116.9 (2C), 116.7, 116.5, 115.5, 114.8; 112.3, 112.1, 105.6, 56.3, 56.2 (2C), 42.2, 29.3.

MS (ESI) m/z: 1017 (M)⁺.

Rf: 0.24 (CH₂Cl₂:MeOH, 40:1).

General procedure K (starting from 167 and n-octanoic acid) and chromatography on silica gel (hexane:EtOAc, 2:1) to afford 90 as a yellow oil (7.8 mg, 90%).

¹H NMR (300 MHz, CDCl₃) δ 7.18-7.15 (m, 1H), 7.08-7.02 (m, 3H), 6.86 (s, 1H), 6.72-6.69 (m, 2H), 4.93-4.89 (m, 1H), 4.76-4.72 (m, 1H), 3.84 (s, 3H), 3.78 (s, 3H), 3.43 (s, 3H), 3.17 (br t, 2H), 2.63-2.47 (m, 6H), 1.78-1.69 (m, 6H), 1.32-1.25 (m, 18H), 0.90-0.88 (m, 15H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 171.7, 171.4, 155.2, 152.2, 151.2, 147.8, 144.9, 142.6, 140.0, 139.0, 138.5, 133.3, 132.7, 127.3, 123.9, 123.1, 123.0, 120.3, 116.0, 115.4, 114.6, 113.3, 112.1, 111.9, 105.5, 56.1, 55.9, 55.8, 42.2, 34.0, 33.9, 33.8, 31.7 (2C), 31.6, 29.2, 29.0, 28.9 (5C), 25.1, 24.9 (2C), 22.6 (3C), 14.1 (3C).

MS (ESI) m/z: 902 (M+23)⁺, 880 (M+1)⁺.

Rf: 0.38 (hexane: EtOAc, 2:1).

A solution of 162 (6.0 mg, 0.01 mmol) was added to a suspension of NaBH₄ (1.0 mg, 0.02 mmol) in anhydrous THF (2 mL) at 0° C. under Argon atmosphere. The reaction mixture was stirred at 23° C. for 3 h, then H₂O (5 mL) was slowly added at 0° C. The mixture was extracted with CH₂Cl₂ (3×10 mL), dried over anhydrous Na₂SO₄, filtered, and evaporated under reduced pressure to give 91 as a white solid (6.0 mg, quant).

¹H NMR (300 MHz, CDCl₃) δ 6.84 (s, 1H), 6.80 (br s, 2H), 6.80 (s, 1H), 6.72 (s, 1H), 6.67 (s, 1H), 6.51 (s, 1H), 6.42 (s, 1H), 4.59 (s, 2H), 4.49-4.44 (m, 2H), 4.28-4.00 (m, 2H), 3.88 (s, 3H), 3.58 (s, 3H), 3.54 (s, 3H), 3.40 (s, 3H), 3.07 (m, 2H), 1.36 (d, J=6.1 Hz, 6H), 1.31 (d, J=6.1 Hz, 6H).

MS (ESI) m/z: 626 (M+23)⁺.

Rf: 0.10 (CH₂Cl₂:MeOH, 20:1).

A suspension of 95 (7.0 mg, 0.0135 mmol), methanesulfonyl chloride (6 mL, 0.077 mmol), pyridine (6 mL, 0.077 mmol) and DMAP (1 mg, 0.008 mmol) in anhydrous CH₂Cl₂ (2 mL) was stirred at 23° C. for 48 h under Argon atmosphere. The solvent was evaporated under reduced pressure and the residue was purified by chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to give 92 as a yellow solid (7.5 mg, 83%).

¹H NMR (300 MHz, CDCl₃) δ 7.50 (d, J=8.1 Hz, 1H), 7.32 (br s, 1H), 7.21 (br d, J=8.1 Hz, 1H), 7.14 (br s, 1H), 6.77 (s, 1H), 6.68 (s, 1H), 6.61 (s, 1H), 4.99-4.94 (m, 1H), 4.70-4.66 (m, 1H), 3.90 (s, 3H), 3.87 (s, 3H), 3.46 (s, 3H), 3.37 (s, 3H), 3.31 (s, 3H), 3.19 (s, 3H), 3.12 (br t, J=6.3 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 154.7, 152.6, 149.4, 148.0, 147.7, 144.8, 138.0, 137.0, 135.9 (2C), 126.7, 125.5, 123.7, 119.3, 117.3, 115.5, 113.7, 111.1, 108.4, 105.7, 56.4, 56.0, 55.8, 55.3, 42.5, 39.1, 38.6, 29.7.

MS (ESI) m/z: 694 (M+23)⁺, 672 (M+1)⁺.

Rf: 0.50 (CH₂Cl₂:MeOH, 20:1).

General procedure F (starting from 95 and hydrocinnamoyl chloride) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 93 as a white solid (4.9 mg, 43%).

¹H NMR (300 MHz, CDCl₃) δ 7.36-7.22 (m, 9H), 7.15-7.02 (m, 3H), 6.76 (s, 1H), 6.70 (s, 1H), 6.68 (s, 1H), 4.90-4.74 (m, 2H), 3.90 (s, 3H), 3.75 (s, 3H), 3.40 (s, 3H), 3.38 (s, 3H), 3.16-2.89 (m, 10H).

MS (ESI) m/z: 802 (M+23)⁺, 780 (M+1)⁺.

Rf: 0.38 (hexane:EtOAc, 1:1).

General procedure L (starting from 167 and Ac₂O) to afford 94 as a brown solid (11.0 mg, 91%.).

¹H NMR (300 MHz, CDCl₃) δ 7.20 (d, J=7.8 Hz, 1H), 7.09-7.04 (m, 3H), 6.87 (s, 1H), 6.70 (br s, 2H), 4.92-4.86 (m, 1H), 4.79-4.72 (m, 1H), 3.85 (s, 3H), 3.79 (s, 3H), 3.43 (s, 3H), 3.17 (t, J=6.3 Hz, 2H), 2.35 (s, 3H), 2.23 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 169.0, 168.8, 168.5, 155.1, 152.1, 151.1, 147.7, 144.8, 139.9, 138.8, 138.4, 134.9, 133.4, 132.9, 127.2, 123.9, 123.1, 120.3, 120.0, 116.1, 115.4, 114.6, 112.1, 111.9, 105.5, 56.1, 56.0, 55.8, 42.1, 29.2, 20.6 (2C), 20.5.

MS (ESI) m/z: 650 (M+23)⁺, 628 (M+1)⁺.

Rf: 0.28 (hexane:EtOAc, 1:1).

General procedure A (starting from 162) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 95 as a yellow solid (170 mg, 61%).

¹H NMR (300 MHz, CDCl₃) δ 7.14-7.07 (m, 2H), 6.07-6.96 (m, 2H), 6.76 (s, 1H), 6.71 (s, 1H), 6.64 (s, 1H), 5.75 (s, 1H), 5.72 (s, 1H), 4.98-4.89 (m, 1H), 4.69-4.59 (m, 1H), 3.89 (s, 3H), 3.87 (s, 3H), 3.50 (s, 3H), 3.38 (s, 3H), 3.15-3.09 (m, 2H).

¹³C NMR (75 MHz, DMSO-d₆) δ 154.3, 148.8, 148.5, 147.0, 146.9, 146.6, 145.7, 144.5, 135.5, 127.7, 126.8, 125.4, 123.4, 119.4, 116.3, 114.7, 112.5, 111.7, 108.7, 105.0, 103.6, 56.0, 55.5, 55.1, 54.5, 42.0, 27.7.

MS (ESI) m/z: 516 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 20:1).

General procedure B (starting from 46) to afford 96.

¹H NMR (300 MHz, CD₃OD) δ 7.60-7.40 (m, 2H), 7.40-7.20 (m, 2H), 7.00-6.80 (m, 2H), 4.77 (br s, 2H), 4.75-4.40 (m, 3H), 3.86 (s, 3H), 3.80 (s, 3H), 3.44 (s, 3H), 3.42 (s, 3H), 3.05 (br s, 2H), 2.00-1.70 (m, 9H).

MS (ESI) m/z: 745 (M+1)⁺.

General procedure E (starting from 46, reaction time 30 h) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 97 as a yellow solid (16 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=7.6 Hz, 1H), 7.40-7.05 (m, 6H), 6.78 (d, J=8.4 Hz, 1H), 5.20-5.00 (m, 3H), 4.80-4.50 (m, 3H), 3.86 (s, 3H), 3.81 (s, 3H), 3.48 (s, 3H), 3.43 (s, 3H), 1.68 (d, J=7.1 Hz, 3H), 1.62 (d, J=7.2 Hz, 3H), 1.55 (d, J=7.1 Hz, 3H), 1.49 (s, 18H), 1.46 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 171.6, 171.3, 155.3, 155.0, 154.9, 153.1, 152.2, 147.6, 145.5, 145.4, 141.6, 140.0, 139.5, 138.8, 134.5, 133.2, 128.3, 128.2, 123.9, 123.6, 123.5, 121.0, 118.3, 115.8, 115.1, 112.1, 109.0, 106.8, 106.2, 104.2, 80.3, 80.1 (2C), 60.8, 56.2, 55.8, 55.6, 49.6, 49.3 (2C), 28.3 (9C), 18.6 (2C), 18.3.

MS (ESI) m/z: 1043 (M)⁺.

Rf: 0.42 (hexane:EtOAc, 1:1).

General procedure E (starting from 141, reaction time 18 h) and chromatography on silica gel (CH₂Cl₂) to afford 98 as a white solid (4 mg, 66%).

¹H NMR (300 MHz, CDCl₃) δ 9.25 (d, J=7.6 Hz, 1H), 8.39 (d, J=1.8 Hz, 2H), 8.32 (d, J=1.8 Hz, 2H), 8.27 (d, J=1.8 Hz, 2H), 8.01 (t, J=1.8 Hz, 1H), 7.97 (t, J=1.8 Hz, 1H), 7.94 (t, J=1.8 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.40-7.25 (m, 3H), 7.19 (s, 1H), 7.09 (d, J=7.6 Hz, 1H), 6.91 (s, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.60 (s, 3H), 3.52 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 162.3, 162.1, 162.0, 154.9, 153.3, 152.4, 147.7, 145.5, 141.9, 140.1, 139.5, 139.4, 139.0, 138.8, 134.8, 133.2, 132.2, 132.1, 132.0, 131.9, 131.8, 130.9, 128.8, 128.3, 124.0, 123.7, 123.6, 123.5, 123.3, 123.2, 121.0, 118.1, 116.1, 115.2, 112.2, 109.1, 106.5, 106.3, 104.5, 61.0, 56.3, 55.9, 55.8. MS (APCI) m/z: 1315 (M)⁺.

Rf: 0.73 (CH₂Cl₂).

General procedure D (starting from 1,2,3,4,5-tetrafluorobenzoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 99 as a white solid (35 mg, 71%).

¹H NMR (300 MHz, CDCl₃) δ 7.90-7.70 (m, 3H), 7.38 (d, J=8.0 Hz, 1H), 7.30-7.15 (m, 3H), 6.76 (s, 2H), 5.00-4.80 (br s, 1H), 4.80-4.60 (br s, 1H), 3.85 (s, 3H), 3.83 (s, 3H), 3.48 (s, 3H), 3.47 (s, 3H), 3.05 (br s, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 159.9, 155.2, 152.3, 152.2, 147.7, 145.2, 141.4, 141.3, 139.7, 138.6, 135.0, 127.2, 124.0, 123.6, 123.0, 119.2, 116.7, 116.0, 115.3, 115.1, 114.2, 113.9, 112.1, 108.3, 105.8, 61.2, 56.6, 56.1, 55.9, 42.1, 22.6.

MS (ESI) m/z: 1059 (M)⁺.

Rf: 0.46 (CH₂Cl₂).

General procedure I (starting from 1) and chromatography on silica gel (CH₂Cl₂) to afford 100 as a white solid (30 mg, 85%).

¹H NMR (300 MHz, CDCl₃) δ 7.49 (d, J=8.6 Hz, 1H), 7.30-7.20 (m, 3H), 6.63 (s, 1H), 6.62 (s, 1H), 5.00-4.90 (m, 1H), 4.80-4.60 (m, 1H), 3.96 (s, 3H), 3.94 (s, 3H), 3.47 (s, 3H), 3.37 (s, 3H), 3.25-3.15 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 154.2, 152.7, 152.0, 147.9, 144.4, 141.7, 140.3, 138.7, 137.2, 136.5, 134.1, 126.1, 123.7, 123.4, 122.4, 120.7, 119.3, 117.8, 116.4, 115.4, 112.1, 109.0, 105.7, 61.4, 56.7, 55.8, 55.5, 41.8, 22.6.

MS (ESI) m/z: 927 (M)⁺.

Rf: 0.57 (CH₂Cl₂).

General procedure G (starting from methanesulfonic acid 5-isopropoxy-6,7-dimethoxy-3-isoquinolin-8-ylmethyl ester) and chromatography on silica gel (hexane:EtOAc, from 3:1 to 1:1) to afford 101 as a white solid (4 mg, 19%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.5 Hz, 1H), 7.38 (d, J=7.5 Hz, 1H), 7.26 (s, 1H), 7.05 (s, 2H), 7.00-6.90 (m, 2H), 4.80-4.50 (m, 3H), 3.95 (s, 5H), 3.85 (s, 3H), 3.75 (s, 3H), 3.54 (s, 2H), 2.92 (s, 3H), 1.50-1.30 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 154.3, 151.0, 147.8, 146.8, 146.6, 146.4, 146.2, 140.7, 140.2, 134.7, 128.9, 128.8, 124.3, 124.0, 123.7, 123.1, 120.2, 116.5, 116.2, 115.4, 109.8, 107.7, 105.7, 103.4, 77.2, 71.7, 71.4, 68.6, 61.6, 60.6, 57.6, 56.0, 55.8, 22.8, 22.0, 21.8.

MS (ESI) m/z: 722 (M-^(i)Pr)⁺.

Rf: 0.50 (hexane:EtOAc, 1:1).

General procedure L (starting from 2 and Ac₂O) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 102 as a white solid (5.4 mg, 96%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.6 Hz, 1H), 7.40-7.00 (m, 6H), 6.81 (s, 1H), 3.89 (s, 3H), 3.83 (s, 3H), 3.48 (s, 3H), 3.45 (s, 3H), 2.49 (s, 3H), 2.37 (s, 3H), 2.32 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 168.8, 168.7, 155.0, 153.2, 152.4, 147.8, 145.5, 141.9, 140.3, 139.8, 139.0, 134.3, 133.3, 128.4, 124.1, 123.6, 123.4, 121.0, 118.2, 115.7, 115.1, 112.2, 112.1, 109.0, 106.6, 106.1, 104.1, 60.8, 56.2, 55.7, 55.6, 20.6 (3C).

MS (ESI) m/z: 678 (M+23)⁺.

Rf: 0.38 (hexane:EtOAc, 1:1).

General procedure L (starting from 1 and Ac₂O) and chromatography on silica gel (hexane:EtOAc, 1:2) to afford 103 as a white solid (12 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.30-7.05 (m, 4H), 6.68 (s, 2H), 4.95-4.80 (m, 1H), 4.80-4.60 (m, 1H), 3.83 (s, 3H), 3.81 (s, 3H), 3.42 (s, 3H), 3.38 (s, 3H), 3.00-2.95 (m, 2H), 2.40 (s, 3H), 2.35 (s, 3H), 2.31 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 168.9, 168.8, 168.7, 155.1, 152.2, 151.8, 147.7, 144.9, 141.6, 141.2, 140.0, 138.9, 134.9, 134.1, 127.2, 123.9, 123.2, 122.6, 119.1, 116.0, 115.8, 114.8, 114.7, 111.9, 107.5, 105.4, 60.8, 56.2, 55.7, 55.4, 41.9, 22.2, 20.6 (2C), 20.5.

MS (ESI) m/z: 680 (M+23)⁺.

Rf: 0.32 (hexane:EtOAc, 1:1).

General procedure H (starting from 6,7-dimethoxy-5-isopropoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (hexane:CH₂Cl₂:Et₂O, from 5:5:1 to 5:5:2) to afford 104 as a white solid (1.58 g, 56%).

¹H NMR (300 MHz, CDCl₃) δ 7.15-7.00 (m, 3H), 6.91 (s, 1H), 6.63 (s, 1H), 6.59 (s, 1H), 4.73 (t, J=7.0 Hz, 2H), 4.65-4.50 (m, 3H), 3.82 (s, 6H), 3.41 (s, 3H), 3.33 (s, 3H), 3.14 (t, J=6.8 Hz, 2H), 1.39 (t, J=6.3 Hz, 12H), 1.31 (d, J=6.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 151.7, 151.3, 148.6, 147.0, 146.9, 146.5, 145.9, 142.5, 135.5, 128.6, 128.1, 123.4, 123.0, 121.1, 116.9, 115.6, 114.6, 113.8, 110.3, 104.9, 104.8, 103.5, 75.7, 71.8, 71.4, 60.5, 56.2, 55.4, 55.1, 42.3, 22.7, 21.9, 21.8.

MS (ESI) m/z: 658 (M+1)⁺.

Rf: 0.56 (hexane:EtOAc, 1:1).

General procedure J (starting from 163, reaction time 22 h) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 105 (10 mg, 56%).

¹H NMR (300 MHz, CDCl₃) δ 7.12-7.03 (m, 3H), 6.92 (s, 1H), 6.81 (s, 1H), 6.69 (s, 1H), 6.67 (s, 1H), 4.85-4.69 (m, 2H), 4.64-4.40 (m, 2H), 3.82 (s, 3H), 3.43 (s, 3H), 3.39 (s, 3H), 3.08 (t, J=6.6 Hz, 2H), 1.42-1.37 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 151.3, 147.0, 146.9, 146.5, 146.0, 145.8, 145.1, 136.0, 129.3, 128.7, 128.2, 127.5, 125.0, 123.4, 119.7, 116.9, 114.6, 114.2, 110.4, 108.3, 104.9, 103.5, 71.8, 71.4, 56.2, 55.5, 55.3, 42.4, 28.5, 21.9 (2C), 21.8 (2C).

MS (ESI) m/z: 608 (M+23)⁺, 586 (M+1)⁺.

Rf: 0.30 (hexane:EtOAc, 1:1).

General procedure D (starting from 109 and cyclohexanepropionic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, from 100:1 to 50:1) to afford 106 as a white solid (33 mg, 72%).

¹H NMR (300 MHz, CDCl₃) δ 7.20 (d, J=7.9 Hz, 1H), 7.14-7.07 (m, 3H), 6.94 (s, 1H), 6.79 (s, 1H), 6.70 (s, 1H), 4.92-4.84 (m, 1H), 4.79-4.70 (m, 1H), 3.80 (s, 3H), 3.42 (s, 3H), 3.35 (s, 3H), 3.11 (t, J=6.7 Hz, 2H), 2.64-2.54 (m, 6H), 1.81-1.60 (m, 21H), 1.39-1.12 (m, 12H), 0.98-0.91 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 172.1, 171.9, 171.8, 155.1, 152.3, 149.9, 147.7, 144.9, 140.1, 139.5, 139.0, 135.1, 133.8, 127.1, 125.9, 125.5, 123.9, 123.1, 122.6, 115.9 (2C), 114.8, 114.6, 111.9, 109.7, 105.4, 56.2, 55.7, 55.5, 42.4, 37.2 (3C), 37.1 (3C), 32.9 (6C), 32.2 (3C), 31.6 (3C), 28.0, 26.5 (3C), 26.2 (3C).

MS (APCI) m/z: 916 (M+1)⁺.

Rf: 0.17 (hexane:EtOAc, 4:1).

General procedure E (starting from 110, reaction time 2 h) and chromatography on silica gel (hexane:EtOAc, 2:1) to afford 107 (283 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.3 Hz, 1H), 7.29-7.17 (m, 2H), 7.12-7.10 (m, 2H), 7.04-7.02 (m, 2H), 6.98 (s, 1H), 6.76 (s, 1H), 4.70-4.56 (m, 3H), 3.84 (s, 3H), 3.44 (s, 3H), 3.43 (s, 3H), 1.44-1.39 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 155.3, 151.2, 150.0, 148.3, 147.7, 147.0, 146.4, 146.3, 134.2, 129.2, 128.6, 124.6, 123.8, 122.9, 118.8, 116.7, 114.9, 112.1, 110.8, 110.2, 109.8, 107.6, 105.5, 105.3, 103.2, 71.6, 71.3, 71.0, 56.0, 55.3, 55.0, 21.8 (3C), 21.7, 21.7, 21.6.

MS (ESI) m/z: 648 (M+23)⁺, 626 (M+1)⁺.

Rf: 0.35 (hexane:EtOAc, 2:1).

General procedure L (starting from 109 and Ac₂O) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 40:1) to afford 108 as a white solid (38 mg, 76%).

¹H NMR (300 MHz, CDCl₃) δ 7.22 (d, J=7.9 Hz, 1H), 7.15-7.09 (m, 3H), 6.95 (s, 1H), 6.79 (s, 1H), 6.69 (s, 1H), 4.92-4.71 (m, 2H), 3.81 (s, 3H), 3.43 (s, 3H), 3.35 (s, 3H), 3.12 (t, J=6.7 Hz, 2H), 2.35 (s, 3H), 2.31 (s, 3H), 2.30 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 169.0, 168.8, 168.6, 155.1, 152.2, 149.9, 147.7, 144.9, 140.0, 139.4, 138.9, 135.0, 134.0, 127.1, 125.9, 125.7, 123.9, 123.2, 122.6, 116.0, 115.9, 114.9, 114.6, 112.0, 109.7, 105.4, 56.2, 55.7, 55.5, 42.5, 28.1, 20.6 (3C).

MS (ESI) m/z: 628 (M+1)⁺.

Rf: 0.32 (CH₂Cl₂:MeOH, 100:1).

General procedure A (starting from 110) and chromatography on silica gel (CH₂Cl₂:MeOH, from 20:1 to 10:1 to 5:1) to afford 109 as a pale brown solid (1.11 g, 97%).

¹H NMR (300 MHz, DMSO-d₆) 59.64 (s, 1H), 9.41 (s, 1H), 9.24 (s, 1H), 7.01 (s, 1H), 6.99 (d, J=8.1 Hz, 1H), 6.88 (d, J=8.4 Hz, 1H), 6.78 (s, 1H), 6.73 (s, 1H), 6.67 (s, 1H), 6.59 (s, 1H), 4.58 (t, J=6.5 Hz, 2H), 3.73 (s, 3H), 3.34 (s, 3H), 3.25 (s, 3H), 2.99 (t, J=6.4 Hz, 2H).

¹³C NMR (75 MHz, DMSO-d₆) δ 154.3, 148.5, 147.1, 146.9, 146.5, 146.0, 145.7, 144.4, 135.9, 127.7, 127.1, 125.5, 123.4, 118.1, 116.3, 115.3, 114.7, 114.3, 112.2, 109.2, 108.8, 105.1, 103.6, 56.0, 55.0, 54.7, 42.0, 27.5.

MS (ESI) m/z: 524 (M+23)⁺.

Rf: 0.55 (CH₂Cl₂:MeOH 10:1).

General procedure H (starting from 6-Isopropoxy-7-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (hexane:CH₂Cl₂:Et₂O, 5:5:2) to afford 110 as a pale yellow solid (1.27 g, 47%).

¹H NMR (300 MHz, CDCl₃) δ 7.08-7.04 (m, 3H), 6.92 (s, 1H), 6.76-6.74 (m, 2H), 6.67 (s, 1H), 4.87-4.71 (m, 2H), 4.65-4.48 (m, 3H), 3.82 (s, 3H), 3.42 (s, 3H), 3.33 (s, 3H), 3.09 (t, J=6.6 Hz, 2H), 1.41-1.36 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 155.5, 151.2, 148.5, 147.2, 146.9, 146.8, 146.4, 145.8, 135.9, 128.5, 128.1, 126.3, 123.3, 120.1, 116.8, 114.8, 114.6, 114.5, 113.6, 110.3, 109.1, 104.8, 103.4, 71.7, 71.3, 71.2, 56.1, 55.4, 55.0, 42.3, 28.5, 22.0 (2C), 21.8, 21.8, 21.7 (2C).

MS (ESI) m/z: 628 (M+1)⁺.

Rf: 0.28 (hexane:CH₂Cl₂:Et₂O, 5:5:2).

General procedure E (starting from 162, reaction time 3 h) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 111 as a white solid (176.3 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 9.25 (d, J=7.3 Hz, 1H), 7.19-7.04 (m, 6H), 6.97 (s, 1H), 6.76 (s, 1H), 4.66-4.56 (m, 2H), 3.99 (s, 3H), 3.84 (s, 3H), 3.47 (s, 3H), 3.45 (s, 3H), 1.44-1.40 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.5, 151.3, 150.0, 149.1, 147.8, 147.1, 146.6, 146.5, 134.3, 129.4, 128.6, 124.7, 123.9, 123.3, 119.1, 116.8, 115.0, 112.2, 111.0, 109.9, 107.8, 107.3, 105.4, 105.3, 103.4, 71.7, 71.4, 56.1, 55.9, 55.9, 55.4, 55.1, 21.9, 21.8.

MS (ESI) m/z: 598 (M+1)⁺.

Rf: 0.50 (hexane:EtOAc, 1:1).

General procedure E (starting from 116, reaction time 23 h) and chromatography on silica gel (hexane:EtOAc, from 2:1 to 1:1) to afford 112 (17 mg, 99%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.11 (d, J=7.5 Hz, 1H), 7.50-7.00 (m, 20H), 6.78 (s, 1H), 6.71 (d, J=7.5 Hz, 1H), 3.80 (s, 3H), 3.78 (s, 3H), 3.47 (s, 3H), 3.40 (s, 3H), 3.20-2.80 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 170.8, 170.7, 155.0, 153.1, 152.3, 147.7, 145.4, 141.8, 140.2, 140.1, 140.0, 139.9, 139.7, 138.9, 134.2, 133.2, 128.7, 128.6, 128.5, 128.4, 128.3, 126.7, 126.5, 126.4, 124.0, 123.3, 123.2, 120.9, 118.2, 115.6, 115.0, 112.1, 108.9, 106.5, 106.1, 104.1, 60.8, 56.2, 55.7, 55.6, 35.5 (3C), 31.0, 30.9, 30.8.

MS (ESI) m/z: 948 (M+23)⁺, 926 (M+1)⁺.

Rf: 0.39 (hexane:EtOAc, 2:1).

General procedure E (starting from 121, reaction time 22 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 80:1) to afford 113 (43 mg, 86%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.3 Hz, 1H), 7.37-7.20 (m, 20H), 7.08-7.03 (m, 2H), 6.80 (d, J=2.2 Hz, 1H), 5.29-5.02 (m, 3H), 4.90-4.88 (m, 3H), 3.85 (s, 3H), 3.44 (s, 6H), 3.41-3.23 (m, 6H), 1.46 (s, 9H), 1.43 (s, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.1, 170.0, 169.9, 155.1 (2C), 154.9, 152.3, 150.9, 147.6, 145.3, 140.5, 139.9, 139.3, 135.8 (2C), 134.5, 133.4, 125.0 (9C), 128.6 (6C), 128.1, 128.0, 127.1 (2C), 124.0, 123.7, 123.6, 123.1, 120.7, 115.8, 115.1, 112.8, 112.3, 112.1, 109.1, 106.4, 106.1, 80.1 (3C), 56.2 (2C), 55.7, 55.6, 55.5, 54.4, 38.1 (3C), 28.2 (9C).

MS (ESI) m/z: 1263 (M+23)⁺, 1241 (M+1)⁺.

Rf: 0.56 (CH₂Cl₂:MeOH, 50:1).

General procedure D (starting from 1 and (L)-N-Boc-Cys(Fm)) and chromatography on silica gel (hexane:EtOAc, 1:1) to afford 114 as a white solid (140 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 7.85-7.65 (m, 12H), 7.45-7.25 (m, 12H), 7.25-7.05 (m, 4H), 6.64 (t, J=2.9 Hz, 2H), 5.50-5.30 (m, 3H), 4.90-4.70 (m, 4H), 4.60 (br s, 1H), 4.25-4.10 (m, 3H), 3.75 (s, 3H), 3.72 (s, 3H), 3.35 (s, 3H), 3.33 (s, 3H), 3.30-3.10 (m, 12H), 2.95 (m, 2H), 1.48 (s, 18H), 1.46 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 169.3, 169.1, 155.1, 154.9, 151.9, 151.7, 147.4, 145.7, 145.5, 144.8, 141.1, 141.0, 140.9, 139.5, 138.4, 134.7, 134.4, 127.7, 127.6, 127.5, 127.0, 126.9, 124.8, 124.7, 124.6, 123.7, 123.2, 122.6, 120.0, 119.9, 119.8, 119.3, 116.2, 115.6, 114.8, 114.7, 111.9, 107.7, 105.4, 80.5, 80.4, 80.3, 60.8, 56.1, 55.6, 55.5, 53.6, 53.4, 46.9, 41.9, 37.3, 37.2, 37.1, 35.6, 35.2, 31.9, 29.6, 28.3, 22.2.

MS (ESI) m/z: 1698 (M+23)⁺, 1676 (M+1)⁺.

Rf: 0.19 (hexane:EtOAc, 2:1).

General procedure B (starting from 121) to afford 115 as a white solid (17 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.44-7.35 (m, 17H), 7.26 (d, J=8.1 Hz, 1H), 7.10 (d, J=1.6 Hz, 1H), 7.06 (s, 1H), 6.88 (d, J=3.5 Hz, 1H), 6.78 (d, J=3.5 Hz, 1H), 4.79-4.70 (m, 3H), 4.63 (t, J=6.7 Hz, 2H), 3.89 (s, 3H), 3.53-3.26 (m, 6H), 3.44 (s, 3H), 3.36 (s, 3H), 3.16 (t, J=6.7 Hz, 2H).

MS (ESI) m/z: 965 (M+23)⁺, 943 (M+1)⁺.

General procedure F (starting from 1 and hydrocinnamoyl chloride) and chromatography on silica gel (hexane:EtOAc, from 2:1 to 1:1) to afford 116 as a white solid (32 mg, 74%).

¹H NMR (300 MHz, CDCl₃) δ 7.40-7.20 (m, 15H), 7.15-7.05 (m, 3H), 7.02 (s, 1H), 6.66 (s, 2H), 4.80-4.70 (m, 1H), 4.70-4.50 (m, 1H), 3.78 (s, 3H), 3.73 (s, 3H), 3.39 (s, 3H), 3.38 (s, 3H), 3.20-2.80 (m, 12H), 2.71 (t, J=6.4 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 170.8, 170.7, 170.6, 155.0, 152.2, 151.8, 147.6, 144.9, 141.5, 141.1, 140.2, 140.1, 140.0, 139.9, 138.9, 134.8, 134.0, 128.6, 128.5, 128.4, 128.3, 128.3, 128.2, 127.1, 126.6, 126.4, 126.3, 123.8, 123.2, 122.5, 119.1, 115.9, 115.7, 114.7, 114.6, 111.8, 107.5, 105.4, 60.7, 56.2, 55.7, 55.5, 41.8, 35.5, 35.4, 35.3, 30.9, 30.8, 30.8, 21.9.

MS (ESI) m/z: 950 (M+23)⁺, 928 (M+1)⁺.

Rf: 0.37 (hexane:EtOAc, 2:1).

General procedure E (starting from 124, reaction time 31 h) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 30:1) to afford 117 as a brownish solid (40 mg, 67%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.3 Hz, 1H), 7.50-7.00 (m, 6H), 6.85-6.70 (m, 1H), 4.80-4.40 (m, 3H), 3.86 (s, 3H), 3.80 (s, 3H), 3.75-3.50 (m, 6H), 3.47 (s, 3H), 3.42 (s, 3H), 2.50-2.20 (m, 6H), 2.20-1.85 (m, 6H), 1.52 (s, 9H), 1.49 (s, 9H), 1.47 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.5, 170.9, 170.8, 155.0, 154.6, 154.4, 154.3, 153.8, 153.7, 152.4, 147.7, 145.5, 141.6, 140.1, 139.7, 134.4, 133.2, 124.2, 123.6, 123.4, 123.2, 121.0, 118.7, 115.7, 115.2, 112.3, 111.9, 107.5, 106.6, 106.1, 104.1, 80.2, 80.1, 79.9, 60.7, 59.0, 58.9, 56.1, 55.8, 55.7, 55.5, 46.6, 46.5, 46.4, 28.5, 28.4, 28.2, 24.5, 24.4, 24.3, 23.6, 23.5, 23.4.

MS (ESI) m/z: 1143 (M+23)⁺.

Rf: 0.32 (CH₂Cl₂:MeOH, 50:1).

General procedure B (starting from 124) to afford 118 as a white solid (25 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.49 (d, J=7.8 Hz, 1H), 7.41 (s, 1H), 7.30-7.20 (m, 2H), 6.81 (d, J=7.9 Hz, 1H), 6.76 (d, J=9.7 Hz, 1H), 4.90-4.70 (m, 5H), 3.87 (s, 3H), 3.81 (s, 3H), 3.60-3.40 (m, 12H), 3.08 (br t, 2H), 2.70-2.30 (m, 6H), 2.30-2.00 (m, 6H).

MS (ESI) m/z: 823 (M+1)⁺.

General procedure B (starting from 125) to afford 119 as a white solid (28 mg, 99%).

¹H NMR (300 MHz, CD₃OD) δ 7.50-7.35 (m, 16H), 7.30-7.20 (m, 2H), 7.15-7.10 (m, 1H), 6.82-6.75 (m, 2H), 4.85-4.55 (m, 5H), 3.88 (s, 3H), 3.81 (s, 3H), 3.60-3.40 (m, 12H), 2.94 (br s, 2H).

MS (ESI) m/z: 973 (M)⁺.

General procedure E (starting from 125, reaction time 31 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 50:1) to afford 120 as a yellow solid (54 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 9.12 (dd, J=7.5, 2.5 Hz, 1H), 7.40-7.20 (m, 19H), 7.10-7.00 (m, 2H), 6.79 (d, J=4.0 Hz, 1H), 5.20-4.80 (m, 6H), 3.86 (s, 3H), 3.85 (s, 3H), 3.48 (s, 3H), 3.44 (s, 3H), 3.40-3.20 (m, 6H), 1.46 (s, 18H), 1.43 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 170.6, 170.0, 169.8, 155.3, 155.1, 155.0, 154.8, 153.1, 152.2, 147.5, 145.3 (2C), 141.7, 139.8, 139.2 (2C), 138.8, 135.8, 135.7, 135.6, 134.6, 133.1 (2C), 129.5 (3C), 129.4 (3C), 128.8 (2C), 128.6 (2C), 128.1, 127.3, 127.2, 123.9, 123.6, 123.3, 120.9, 118.2, 115.8, 115.1, 112.0, 108.9, 106.9, 106.1, 104.2, 80.4, 80.2, 80.0, 60.8, 56.2, 56.1, 55.7, 54.7, 54.3 (2C), 38.1 (2C), 37.8, 28.2 (9C).

MS (ESI) m/z: 1293 (M+23)⁺.

Rf: 0.32 (CH₂Cl₂:MeOH, 60:1).

General procedure D (starting from 109 and Boc-L-Phe-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 121 as a white solid (87 mg, 68%).

¹H NMR (300 MHz, CDCl₃) δ 7.34-7.26 (m, 15H), 7.16 (br s, 2H), 7.10 (s, 1H), 7.05 (s, 1H), 6.91 (s, 1H), 6.78-6.68 (m, 2H), 4.99 (t, J=8.6 Hz, 2H), 4.88-4.72 (m, 6H), 3.81 (s, 3H), 3.41 (s, 3H), 3.34 (s, 3H), 3.30-3.12 (m, 8H), 1.44 (s, 9H), 1.43 (s, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.1, 169.9 (2C), 155.0, 154.9, 152.0, 149.7, 147.5, 144.7, 139.6, 139.0, 138.4, 135.8 (2C), 134.8, 134.2, 129.4 (9C), 129.2, 128.5 (6C), 127.1 (2C), 126.9, 125.9, 125.7, 123.8, 123.1, 122.5, 116.1, 115.8, 114.9, 114.7, 111.8, 109.7, 105.4, 80.0 (3C), 56.1, 55.6, 55.4, 54.3 (3C), 42.3, 38.0 (3C), 28.2 (9C), 27.9.

MS (ESI) m/z: 1265 (M+23)⁺.

Rf: 0.65 (CH₂Cl₂:MeOH, 30:1).

General procedure E (starting from 134, reaction time 22 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 50:1) to afford 122 (47 mg, 94%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.26 (d, J=7.3 Hz, 1H), 7.42 (s, 1H), 7.31-7.26 (m, 2H), 7.23-7.18 (m, 3H), 7.09 (d, J=7.5 Hz, 1H), 6.80 (d, J=7.5 Hz, 1H), 5.09-5.06 (m, 3H), 4.57-4.50 (m, 3H), 3.80 (s, 3H), 3.43 (s, 6H), 2.45-2.34 (m, 3H), 1.50 (s, 9H), 1.47 (s, 18H), 1.14-1.00 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4 (b), 155.7, 154.9, 152.3, 150.9, 147.6, 145.4, 140.6, 140.0, 139.4, 134.5, 133.4, 128.1, 124.0, 123.8, 123.6, 123.1, 120.8, 115.8, 115.1, 112.8, 112.2, 112.2, 109.1, 106.4, 106.1, 80.0 (3C), 58.5 (2C), 56.0, 55.6, 55.5, 55.4, 30.0 (3C), 28.3 (9C), 19.2, 19.1 (2C), 17.2, 17.1 (2C).

MS (ESI) m/z: 1119 (M+23)⁺, 1097 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 100:1).

General procedure E (starting from 139, reaction time 7 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 123 (15 mg, 75%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.5 Hz, 1H), 7.84-7.75 (m, 3H), 7.54 (s, 1H), 7.45 (d, J=8.4 Hz, 1H), 7.37-7.35 (m, 2H), 7.30 (s, 1H), 7.26 (s, 1H), 7.08 (d, J=7.3 Hz, 1H), 6.90 (s, 1H), 3.87 (s, 3H), 3.52 (s, 3H), 3.52 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 159.7 (3C), 154.8, 152.3, 150.8, 148.2, 147.6, 145.4, 140.3, 139.8, 139.2, 135.0, 133.4, 128.0, 124.0, 123.9, 123.8, 123.7, 123.2, 120.7, 116.2, 115.3, 113.8, 113.6, 112.8, 112.4, 112.1, 109.2, 106.6, 106.3, 56.4, 55.9, 55.8.

MS (ESI) m/z: 1050 (M+23)⁺, 1028 (M+1)⁺.

Rf: 0.63 (CH₂Cl₂).

General procedure D (starting from 1 and (L)-N-Boc-Pro) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 20:1) to give 124 as a white solid (105 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.20-7.10 (m, 2H), 7.10-7.00 (m, 2H), 6.70-6.60 (m, 2H), 4.90 (br s, 1H), 4.70-4.40 (m, 4H), 3.78 (s, 6H), 3.70-3.40 (m, 6H), 3.39 (s, 3H), 3.36 (s, 3H), 3.20-2.95 (m, 2H), 2.50-2.20 (m, 6H), 2.15-1.85 (m, 6H), 1.48 (s, 18H), 1.46 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 171.3, 170.9, 170.7, 155.0, 154.4, 153.8, 152.2, 151.7, 147.6, 144.8, 141.4, 141.0, 139.8, 138.7, 134.3, 127.1, 124.0, 123.4, 123.2, 122.6, 119.9, 119.0, 116.0, 115.7, 114.7, 112.1, 111.6, 107.5, 105.4, 80.2, 80.0, 79.9, 60.6, 58.9, 58.8, 56.1, 55.7, 55.6, 55.5, 46.6, 46.5, 46.4, 42.0, 28.3, 24.4, 24.3, 23.6, 23.5, 23.4, 23.3, 22.0.

MS (ESI) m/z: 1145 (M+23)⁺, 1124 (M+1)⁺.

Rf: 0.64 (CH₂Cl₂:MeOH, 20:1).

General procedure D (starting from 1 and (L)-Boc-Phe) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 30:1) to afford 125 as a brown solid (119 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.50-7.25 (m, 15H), 7.20-7.10 (m, 3H), 7.03 (s, 1H), 5.10-5.00 (m, 3H), 5.00-4.80 (m, 3H), 4.75-4.50 (m, 2H), 3.83 (s, 3H), 3.82 (s, 3H), 3.78 (s, 6H), 3.40 (s, 3H), 3.37 (s, 3H), 3.35-3.00 (m, 6H), 2.95-2.85 (m, 2H), 1.44 (s, 9H), 1.43 (s, 9H), 1.42 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4, 169.9 (2C), 155.2, 154.9, 152.1, 151.8, 147.5, 144.8, 141.3, 141.0, 139.6, 138.4, 135.8, 135.6, 134.7, 134.3, 129.5 (3C), 129.4 (2C), 129.2 (2C), 128.7 (2C), 128.5 (2C), 128.4 (2C), 127.2, 127.1, 126.9, 123.7, 123.2, 122.6, 119.4, 116.2, 115.6, 114.8, 114.7, 111.8, 107.6, 105.4, 80.3, 80.1, 80.0, 60.7, 56.1, 55.6, 55.5, 54.3, 53.4, 52.1, 41.8, 38.1 (2C), 37.8, 28.2 (9C), 22.0.

MS (ESI) m/z: 1295 (M+23)⁺, 1273 (M+1)⁺.

Rf: 0.21 (CH₂Cl₂:MeOH, 50:1).

General procedure I (starting from 26) and chromatography on silica gel (CH₂Cl₂) to afford 126 as a pale yellow solid (24.2 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.16 (d, J=7.3 Hz, 1H), 7.54 (d, J=8.1 Hz, 1H), 7.40-7.25 (m, 2H), 7.20 (s, 1H), 7.11 (br s, 2H), 6.98 (s, 1H), 6.76 (s, 1H), 3.99 (s, 3H), 3.98 (s, 3H), 3.50 (s, 3H), 3.46 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 154.4, 152.6, 150.7, 149.8, 147.9, 144.9, 138.8, 137.7, 137.4, 134.3, 129.6, 127.5, 127.2, 125.0, 124.1, 123.7, 122.9, 118.6 (2C, t, J_(C-F)=172.7, 164.5 Hz), 116.3, 113.7, 112.0, 110.2, 107.7, 106.5, 104.7, 56.7, 56.0, 55.8, 55.2.

MS (ESI) m/z: 778 (M+1)⁺.

Rf: 0.36 (CH₂Cl₂).

General procedure E (starting from 140, reaction time 17 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 30:1) to afford 127 (30 mg, 67%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.27-9.23 (m, 1H), 7.47-7.36 (m, 1H), 7.26-7.08 (m, 6H), 6.84-6.78 (m, 1H), 4.56-4.49 (m, 3H), 3.80 (s, 3H), 3.66-3.47 (m, 6H), 3.43 (s, 6H), 2.40-2.29 (m, 6H), 2.04-1.98 (m, 6H), 1.49 (s, 27H).

MS (ESI) m/z: 1091 (M+1)⁺.

Rf: 0.31 (CH₂Cl₂:MeOH 30:1).

General procedure B (starting from 131) to afford 128 as a white solid (25 mg, 99%).

¹H NMR (300 MHz, CD₃OD) δ 7.50-7.40 (m, 2H), 7.30-7.20 (m, 2H), 6.82 (d, J=5.4 Hz, 1H), 6.79 (d, J=7.9 Hz, 1H), 4.90-4.70 (m, 2H), 4.44 (d, J=3.4 Hz, 1H), 4.32 (dd, J=4.2, 1.6 Hz, 1H), 4.24 (d, J=4.3 Hz, 1H), 3.86 (s, 3H), 3.81 (s, 3H), 3.44 (s, 3H), 3.43 (s, 3H), 3.15-3.00 (m, 2H), 2.65-2.40 (m, 3H), 1.30-1.15 (m, 18H).

MS (ESI) m/z: 851 (M+23)⁺, 829 (M+1)⁺.

General procedure E (starting from 131, reaction time 24 h) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 30:1) to afford 129 as a yellow solid (53 mg, 79%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.6 Hz, 1H), 7.40-7.05 (m, 6H), 6.78 (d, J=9.1 Hz, 1H), 5.15-5.05 (m, 3H), 4.65-4.50 (m, 3H), 3.86 (s, 3H), 3.81 (s, 3H), 3.48 (s, 3H), 3.42 (s, 3H), 2.50-2.30 (m, 3H), 1.49 (s, 18H), 1.46 (s, 9H), 1.25-0.95 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.2 (3C), 155.9, 155.7, 154.9, 153.1, 152.2, 147.6, 145.4, 145.3, 141.7, 139.9, 139.4, 138.7, 134.5, 133.1, 128.3 (2C), 124.0, 123.6, 123.5, 121.0, 118.3, 115.8, 115.1, 112.1, 109.0, 106.9, 106.1, 104.2, 80.3, 80.0 (2C), 60.7, 59.0, 58.6 (2C), 56.0, 55.7, 55.6, 31.3, 31.1, 30.9, 28.3 (9C), 19.3, 19.2, 19.0, 17.5, 17.2, 17.1.

MS (ESI) m/z: 1149 (M+23)⁺.

Rf: 0.19 (CH₂Cl₂:MeOH, 50:1).

General procedure E (starting from 136, reaction time 3 d) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 30:1) to afford 130 as a brownish solid (105 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 9.15 (d, J=7.6 Hz, 1H), 7.59 (d, J=8.0 Hz, 1H), 7.37 (d, J=7.5 Hz, 1H), 7.35-7.25 (m, 3H), 7.08 (s, 1H), 6.75 (s, 1H), 3.99 (s, 3H), 3.96 (s, 3H), 3.49 (s, 3H), 3.47 (s, 3H), 3.42 (s, 3H), 3.37 (s, 3H), 3.19 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 154.3, 153.1, 152.9, 148.0, 145.1, 142.6, 138.3, 138.2, 137.7, 135.7, 132.6, 127.6, 125.8, 123.8, 123.6, 121.0, 119.8, 116.6, 115.7, 113.7, 111.8, 109.1, 107.7, 106.4, 104.9, 61.5, 56.6, 55.8, 55.6, 39.7, 39.3, 38.6.

MS (ESI) m/z: 764 (M+1)⁺.

Rf: 0.54 (CH₂Cl₂:MeOH, 50:1).

General procedure D (starting from 1 and (L)-Boc-Valine) and chromatography on silica gel (CH₂Cl₂:MeOH, from 50:1 to 30:1) to afford 131 as a yellow solid (105 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.30-7.10 (m, 2H), 7.08 (s, 2H), 6.63 (t, J=8.9 Hz, 2H), 5.30-5.10 (m, 3H), 4.70 (br s, 1H), 4.66 (br s, 1H), 4.60-4.45 (m, 3H), 3.78 (s, 3H), 3.77 (s, 3H), 3.38 (s, 3H), 3.37 (s, 3H), 3.00 (br t, 2H), 2.45-2.30 (m, 3H), 1.48 (s, 9H), 1.47 (s, 9H), 1.45 (s, 9H), 1.15-0.95 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.6, 170.4 (2C), 155.7, 155.6, 154.9, 152.0, 151.8, 147.4, 144.8, 141.2, 141.0, 139.6, 138.5, 134.7, 134.3, 127.0, 126.9, 123.8, 123.2, 122.6, 119.4, 116.1, 115.6, 114.8, 114.7, 111.8, 107.6, 105.4, 80.1, 80.0, 79.9, 60.6, 58.8, 58.5, 58.4, 56.0, 55.6, 55.4, 41.8, 31.3, 31.1, 30.8, 28.3 (9C), 22.2, 19.2, 19.1, 19.0, 17.4, 17.1, 17.0.

MS (ESI) m/z: 1151.7 (M+23)⁺, 1129.8 (M+1)⁺.

Rf: 0.70 (CH₂Cl₂:MeOH, 20:1).

General procedure B (starting from 131) to afford 132 as a brownish solid (50 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.90-7.40 (m, 19H), 6.80-6.70 (m, 2H), 4.90-4.50 (m, 5H), 3.90 (s, 3H), 3.79 (s, 3H), 3.60-3.30 (m, 12H), 2.72 (br s, 2H).

MS (ESI) m/z: 1090 (M)⁺.

General procedure D (starting from 109 and 3,5-dibromobenzoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, from 200:1 to 100:1) to afford 133 as a white solid (43 mg, 67%).

¹H NMR (300 MHz, CDCl₃) δ 8.28 (d, J=1.8 Hz, 2H), 8.26 (d, J=1.8 Hz, 2H), 8.24 (d, J=1.8 Hz, 2H), 7.95-7.92 (m, 3H), 7.34 (d, J=8.1 Hz, 1H), 7.26 (s, 1H), 7.24-7.20 (m, 2H), 7.08 (s, 1H), 6.88 (s, 1H), 6.81 (s, 1H), 4.94-4.89 (m, 1H), 4.81-4.77 (m, 1H), 3.83 (s, 3H), 3.49 (s, 3H), 3.43 (s, 3H), 3.17 (t, J=6.7 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 169.9, 154.9, 151.9, 149.1, 147.6, 147.4, 144.7, 139.4, 138.3, 135.8, 134.4, 129.4, 129.2, 128.5, 127.1, 126.4, 126.3, 123.6, 123.3, 119.5, 116.3, 114.9, 114.6, 114.4, 111.7, 110.9, 108.4, 105.4, 79.9, 79.9 (2C), 56.0, 55.8, 55.6, 55.4, 54.3 (2C), 42.4, 38.0 (2C), 28.4, 28.2 (6C).

MS (APCI) m/z: 1288 (M+1)⁺.

Rf: 0.72 (CH₂Cl₂).

General procedure B (starting from 140) to afford 134 as a white solid (17 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.49-7.44 (m, 2H), 7.29-7.17 (m, 3H), 6.88-6.75 (m, 2H), 4.79-4.68 (m, 3H), 3.89 (s, 3H), 3.52-3.38 (m, 12H), 3.15 (br t, 2H), 2.63-2.35 (m, 6H), 2.25-2.15 (m, 6H).

MS (ESI) m/z: 793 (M+1)⁺.

General procedure B (starting from 144) to afford 135 as a white solid (16 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.47-7.44 (m, 2H), 7.28 (d, J=8.1 Hz, 1H), 7.19 (s, 1H), 7.15 (s, 1H), 6.90-6.78 (m, 2H), 4.77 (br t, 2H), 4.32 (s, 1H), 4.24 (s, 2H), 3.87 (s, 3H), 3.45 (s, 3H), 3.37 (s, 3H), 3.17 (br t, 2H), 2.54-2.46 (m, 3H), 1.26-1.17 (m, 18H).

MS (ESI) m/z: 799 (M+1)⁺.

To a solution of 1 (25 mg, 0.047 mmol) in anhydrous CH₂Cl₂ (2 mL) under Argon at 0° C., Et₃N (39 μL, 0.28 mmol) and methanesulfonyl chloride (22 μL, 0.28 mmol) were added. The resulting mixture was stirred at 23° C. for 2 h, then quenched with H₂O and extracted with CH₂Cl₂ (3×20 mL).

The combined organic phases were washed with saturated aqueous solution of NaHCO₃, dried over anhydrous Na₂SO₄, filtered, and evaporated under vacuum. The resulting residue was purified on silica gel (CH₂Cl₂:MeOH, 50:1) to afford 136 as a brownish solid (35 mg, 97%).

¹H NMR (300 MHz, CDCl₃) δ 7.51 (d, J=8.0 Hz, 1H), 7.30-7.15 (m, 3H), 6.66 (s, 1H), 6.63 (s, 1H), 5.00-4.90 (m, 1H), 4.75-4.50 (m, 1H), 3.92 (s, 3H), 3.89 (s, 3H), 3.45 (s, 3H), 3.37 (s, 3H), 3.35 (s, 3H), 3.32 (s, 3H), 3.30-3.20 (m, 2H), 3.17 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 154.5, 152.8, 151.9, 148.0, 144.7, 141.8, 141.0, 138.0, 137.0, 135.6, 134.4, 126.3, 125.6, 123.5, 123.1, 121.7, 117.0, 115.5, 115.3, 113.6, 108.2, 105.7, 61.3, 56.5, 55.8, 55.5, 42.0, 39.4, 39.2, 38.6, 23.3. MS (APCI) m/z: 766 (M+1)⁺.

Rf: 0.54 (CH₂Cl₂:MeOH, 50:1).

General procedure D (starting from 1 and (L)-N-Boc-Trp) and chromatography on silica gel (CH₂Cl₂:MeOH, from 30:1 to 20:1) to afford 137 as a brown solid (130 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 8.51 (s, 2H), 8.42 (br s, 1H), 7.70-7.60 (m, 3H), 7.45-7.30 (m, 3H), 7.30-6.95 (m, 9H), 6.87 (s, 1H), 6.70-6.55 (m, 2H), 5.30-5.15 (m, 2H), 5.10-4.90 (m, 3H), 4.80-4.40 (m, 2H), 3.76 (s, 6H), 3.60-3.30 (m, 12H), 2.70 (br s, 2H), 1.45 (s, 27H).

¹³C NMR (75 MHz, CDCl₃) δ 170.8, 170.4 (2C), 155.2, 154.9, 152.0, 151.7, 147.5, 144.6, 141.4, 141.0, 139.6, 138.5, 136.1 (2C), 134.7, 134.1, 127.7, 127.6, 126.8, 123.8, 123.4, 123.1 (2C), 122.6, 122.2, 119.7, 119.6 (2C), 118.7, 118.6, 116.0, 115.6, 114.6, 111.7, 111.3 (2C), 109.8, 109.4, 107.6, 105.4, 80.2, 80.0 (2C), 60.8, 56.1, 55.6, 55.5, 54.4 (2C), 53.4, 41.7, 28.2 (9C+3C), 21.7.

MS (ESI) m/z: 1412 (M+23)⁺, 1391 (M+1)⁺.

Rf: 0.22 (CH₂Cl₂:MeOH, 30:1).

General procedure E (starting from 149, reaction time 2 d) and chromatography on silica gel (CH₂Cl₂:MeOH, from 100:1 to 50:1) to afford 138 as a white solid (29 mg, 83%).

¹H NMR (300 MHz, CDCl₃) δ 9.17 (d, J=7.6 Hz, 1H), 8.85 (d, J=4.5 Hz, 2H), 8.78 (s, 1H), 7.80-7.60 (m, 6H), 7.55-7.20 (m, 11H), 7.18 (s, 1H), 6.89 (s, 1H), 3.94 (s, 3H), 3.90 (s, 3H), 3.60 (s, 3H), 3.53 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 161.2, 160.6, 160.3, 156.4, 156.2, 155.5, 155.4, 155.3, 154.8, 153.3, 152.4, 150.5, 150.4, 150.1, 147.7, 145.4, 141.8, 139.9, 139.4, 138.7, 135.1, 135.0, 134.9, 134.7, 133.2, 129.9, 129.8, 129.7, 128.2, 125.1, 125.0, 124.9, 124.1, 123.7, 123.4, 120.9, 118.2, 117.8, 117.7, 117.7, 116.9, 116.7, 116.5, 115.9, 115.4, 112.3, 112.1, 109.0, 106.9, 106.3, 104.4, 61.0, 56.4, 56.1, 55.9.

MS (ESI) m/z: 1046 (M+1)⁺.

Rf: 0.50 (CH₂Cl₂:MeOH, 50:1).

General procedure D (starting from 1 and 2,3,4,5-tetrafluorobenzoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 139 as a white solid (33 mg, 64%).

¹H NMR (300 MHz, CDCl₃) δ 7.81-7.74 (m, 3H), 7.36 (d, J=8.1 Hz, 1H), 7.25 (s, 1H), 7.23 (s, 1H), 7.19 (s, 1H), 7.10 (s, 1H), 6.86 (s, 1H), 6.78 (s, 1H), 4.97-4.91 (m, 1H), 4.82-4.77 (m, 1H), 3.83 (s, 3H), 3.48 (s, 3H), 3.41 (s, 3H), 3.17 (t, J=6.5 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 159.6 (3C), 154.9, 152.1, 149.7, 147.5, 144.9, 139.5, 138.9, 138.3, 134.9, 134.7, 126.9, 126.2, 126.1, 123.7, 123.3, 122.5, 116.5, 115.9, 115.2, 114.9, 113.8, 113.5, 111.9, 109.9, 105.6, 56.3, 55.9, 55.7, 42.5, 28.1. MS (APCI) m/z: 1030 (M+1)⁺.

Rf: 0.50 (CH₂Cl₂:MeOH, 200:1).

General procedure D (starting from 109 and Boc-L-Pro-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, from 100:1 to 50:1) to afford 140 as a white solid (74 mg, 68%).

¹H NMR (300 MHz, CDCl₃) δ 7.16-7.13 (m, 2H), 7.06-7.03 (m, 2H), 6.90 (s, 1H), 6.78-6.63 (m, 2H), 4.95-4.62 (m, 2H), 4.56-4.46 (m, 3H), 3.78 (s, 3H), 3.69-3.43 (m, 6H), 3.40 (s, 3H), 3.33 (s, 3H), 3.11 (br t, 2H), 2.40-2.26 (m, 6H), 2.08-1.90 (m, 6H), 1.47 (s, 27H).

¹³C NMR (75 MHz, CDCl₃) δ 171.0, 170.8, 170.7, 155.0, 154.3, 153.7, 153.7 (2C), 152.1, 149.7, 147.6, 144.8, 139.8, 139.3, 138.7, 134.1, 125.9, 125.6, 124.0, 123.4, 123.1 (2C), 122.7, 122.2, 116.0, 114.7, 111.6, 109.6, 105.3, 80.1, 80.0, 79.8, 58.9 (2C), 56.1, 55.7, 55.6, 55.5, 46.5, 46.3 (2C), 42.4, 31.5, 30.9, 29.9, 28.3 (9C), 28.0, 24.2, 23.4, 22.5.

MS (ESI) m/z: 1115 (M+23)⁺, 1093 (M+1)⁺.

Rf: 0.18 (CH₂Cl₂:MeOH, 50:1).

General procedure D (starting from 1 and 3,5-dibromobenzoic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 141 as a white solid (61 mg, 98%).

¹H NMR (300 MHz, CDCl₃) δ 8.32 (d, J=1.7 Hz, 2H), 8.30 (d, J=1.8 Hz, 2H), 8.26 (d, J=1.7 Hz, 2H), 7.98 (t, J=1.7 Hz, 1H), 7.96 (t, J=1.7 Hz, 1H), 7.93 (t, J=1.8 Hz, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.25-7.19 (m, 3H), 6.78 (s, 1H), 6.77 (s, 1H), 5.00-4.60 (br s, 2H), 3.84 (s, 3H), 3.82 (s, 3H), 3.48 (s, 6H), 3.10-3.00 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 162.3, 152.5, 152.2, 147.9, 145.2, 141.6, 140.0, 139.5, 139.2, 138.9, 135.1, 134.8, 132.5, 132.2, 129.1, 127.3, 124.0, 123.7, 123.6, 123.5, 123.4, 123.0, 119.3, 116.7, 116.0, 115.1, 112.2, 108.1, 105.8, 61.2, 56.5, 56.1, 55.9, 42.2, 22.6. MS (APCI) m/z: 1319 (M+1)⁺.

Rf: 0.58 (CH₂Cl₂).

General procedure K (starting from 1 and 4-fluorenecarboxylic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, from 200:1 to 100:1) to afford 142 as a white solid (36 mg, 69%).

¹H NMR (300 MHz, CDCl₃) δ 8.60-8.40 (m, 3H), 8.30-8.10 (m, 3H), 7.90-7.70 (m, 3H), 7.65-7.55 (m, 3H), 7.55-7.25 (m, 13H), 6.92 (s, 1H), 6.91 (s, 1H), 5.10-4.70 (br s, 2H), 4.00 (s, 2H), 3.99 (s, 2H), 3.96 (s, 2H), 3.93 (s, 3H), 3.91 (s, 3H), 3.60 (s, 3H), 3.58 (s, 3H), 3.18 (br s, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 166.0, 165.9, 155.1, 152.6, 152.1, 148.0, 145.4. 145.2, 145.1, 145.0, 144.3, 144.2, 144.1, 141.9, 141.8, 141.5, 141.3, 140.4, 140.0, 139.9, 139.8, 139.2, 135.0, 134.3, 129.5, 129.4, 129.3, 129.0, 127.8, 127.7, 127.6, 127.3, 127.0, 126.8, 126.7, 126.2, 126.1, 126.0, 125.4, 125.3, 125.1, 125.0, 124.7, 124.5, 124.1, 123.5, 122.9, 119.6, 116.2, 116.0, 114.9, 112.1, 107.7, 105.6, 61.0, 56.3, 55.8, 55.7, 42.0, 37.0 (3C), 22.5. MS (APCI) m/z: 1108 (M)⁺.

Rf: 0.34 (CH₂Cl₂).

General procedure E (starting from 99, reaction time 63 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 143 as a white solid (27 mg, 93%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.5 Hz, 1H), 7.95-7.70 (m, 3H), 7.46 (d, J=8.2 Hz, 1H), 7.40-7.35 (m, 2H), 7.25 (s, 1H), 7.18 (s, 1H), 7.11 (d, J=7.6 Hz, 1H), 6.88 (s, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 3.58 (s, 3H), 3.51 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 154.7, 153.3, 152.3, 147.6, 145.4, 143.1, 141.7, 139.7, 139.2, 138.5, 135.0, 133.1, 128.2, 123.9, 123.7, 123.6, 120.9, 117.9, 116.1, 115.3, 113.8, 112.1, 109.1, 106.4, 106.3, 104.6, 61.0, 56.4, 55.9, 55.7.

MS (APCI) m/z: 1058 (M+1)⁺.

Rf: 0.54 (CH₂Cl₂).

General procedure D (starting from 109 and Boc-L-Val-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, from 100:1 to 50:1) to afford 144 as a white solid (87 mg, 79%).

¹H NMR (300 MHz, CDCl₃) δ 7.26-7.13 (m, 2H), 7.09 (s, 2H), 6.96 (s, 1H), 6.76 (d, J=7.5 Hz, 1H), 6.68 (d, J=9.5 Hz, 1H), 5.08-5.05 (m, 3H), 4.91-4.69 (m, 2H), 4.52-4.46 (m, 3H), 3.77 (s, 3H), 3.40 (s, 3H), 3.32 (s, 3H), 3.18 (br t, 2H), 2.42-2.38 (m, 3H), 1.48 (s, 9H), 1.45 (s, 18H), 1.11-0.98 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.3 (3C), 155.6, 154.9, 152.1, 149.7, 147.5, 144.8, 139.7, 139.1, 138.5, 134.9, 134.2, 126.9, 126.0, 125.7, 123.8, 123.1, 122.5, 116.1, 115.8, 115.0, 114.6, 111.9, 109.6, 105.4, 79.9 (3C), 58.5, 56.0, 55.6, 55.3, 55.3, 53.4, 42.4, 31.2 (3C), 28.3 (9C), 28.0, 19.1 (2C), 17.1 (4C).

MS (ESI) m/z: 1121 (M+23)⁺, 1099 (M+1)⁺.

Rf: 0.35 (CH₂Cl₂:MeOH, 50:1).

General procedure E (starting from 148, reaction time 24 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 145 as a white solid (27 mg, 87%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.5 Hz, 1H), 7.35-7.00 (m, 6H), 6.81 (s, 1H), 3.87 (s, 3H), 3.82 (s, 3H), 3.49 (s, 3H), 3.45 (s, 3H), 2.76 (t, J=7.6 Hz, 2H), 2.70-2.55 (m, 4H), 1.90-1.60 (m, 18H), 1.50-1.10 (m, 15H), 1.05-0.80 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 172.0, 171.9, 171.8, 155.0, 153.2, 152.4, 147.8, 145.5, 141.8, 140.4, 139.9, 139.1, 134.1, 133.3, 128.4, 124.1, 123.6, 123.3, 121.0, 118.3, 116.5, 115.5, 115.0, 112.2, 108.9, 106.6, 106.1, 104.0, 60.8, 56.2, 55.7, 55.6, 37.2, 37.1, 37.0, 33.0 (4C), 32.9 (4C), 32.4, 32.3, 32.2, 31.6, 31.5, 26.5, 26.2 (6C).

MS (ESI) m/z: 944 (M)⁺.

Rf: 0.35 (CH₂Cl₂).

General procedure E (starting from 153, reaction time 35 h) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 146 (38 mg, 73%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.3 Hz, 1H), 7.42 (s, 1H), 7.32 (d, J=7.9 Hz, 1H), 7.25-7.17 (m, 4H), 7.05 (d, J=7.7 Hz, 1H), 6.79 (d, J=5.9 Hz, 1H), 4.98-4.96 (m, 3H), 4.62-4.56 (m, 3H), 3.80 (s, 3H), 3.44 (s, 3H), 3.43 (s, 3H), 1.87-1.64 (m, 9H), 1.49 (s, 9H), 1.46 (s, 18H), 1.06-0.99 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4 (4C), 155.3, 155.0, 152.3, 150.9, 147.7, 145.4, 140.7, 140.1, 139.6, 134.4, 128.1, 124.0, 123.8, 123.6, 123.2, 120.7, 115.8, 115.1, 112.8, 112.3, 112.2, 109.1, 106.4, 106.2, 80.0 (3C), 56.2 (2C), 55.8 (2C), 55.7, 52.2, 41.7 (2C), 41.5, 28.3 (9C), 24.8 (3C), 23.0, 22.9 (3C), 21.9.

MS (ESI) m/z: 1161 (M+23)⁺, 1139 (M+1)⁺.

Rf: 0.45 (CH₂Cl₂:MeOH, 50:1).

General procedure E (starting from 152, reaction time 40 h) and chromatography on silica gel (CH₂Cl₂:MeOH, from 200:1 to 100:1) to afford 147 (17 mg, 65%) as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.3 Hz, 1H), 7.58-7.50 (m, 6H), 7.31 (s, 1H), 7.21-7.16 (m, 4H), 7.10-7.01 (m, 8H), 6.75 (s, 1H), 3.87 (s, 2H), 3.84 (s, 2H), 3.81 (s, 2H), 3.78 (s, 3H), 3.37 (s, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 167.6, 167.5 (2C), 162.6 (d, J_(C-F)=248.3, 3C), 154.9, 152.3, 150.8, 147.6, 145.4, 140.6, 140.0, 139.5, 134.6, 133.9, 133.8, 133.8, 133.7, 133.6, 133.4, 129.5, 129.3, 128.1, 123.8, 123.6, 123.2, 120.5, 116.4 (6C), 116.1 (6C), 115.9, 115.1, 112.8, 112.3, 112.0, 109.1, 106.4, 106.2.

MS (ESI) m/z: 1026 (M+23)⁺, 1004 (M+1)⁺.

Rf: 0.35 (CH₂Cl₂).

General procedure D (starting from 1 and 3-cyclohexylpropionic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to afford 148 as a white solid (43 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 7.30-7.05 (m, 4H), 6.68 (s, 2H), 5.00-4.80 (m, 1H), 4.80-4.70 (m, 1H), 3.82 (s, 3H), 3.80 (s, 3H), 3.42 (s, 3H), 3.39 (s, 3H), 3.00-2.90 (m, 2H), 2.70-2.50 (m, 6H), 1.90-1.60 (m, 21H), 1.50-1.10 (m, 12H), 1.05-0.90 (m, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 171.9, 171.8, 155.1, 152.3, 151.8, 147.7, 144.9, 141.6, 141.2, 140.1, 139.1, 134.9, 133.9, 127.2, 123.9, 123.2, 122.6, 119.2, 115.9, 115.8, 114.7, 114.6, 111.9, 107.4, 105.4, 60.7, 56.2, 55.7, 55.5, 41.9, 37.2, 37.1, 37.0, 33.0 (4C), 32.9 (4C), 32.4, 32.3, 32.2, 31.6, 31.5, 31.4, 26.5, 26.3 (2C), 26.2 (2C), 26.1(2C), 22.2.

MS (APCI) m/z: 946 (M)⁺.

Rf: 0.37 (CH₂Cl₂:MeOH, 100:1).

General procedure D (starting from 1 and coumarin-3-carboxylic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 50:1) to afford 149 as a white solid (49 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 8.82 (s, 1H), 8.80 (s, 1H), 8.77 (s, 1H), 7.80-7.60 (m, 6H), 7.25-7.15 (m, 7H), 7.15-7.05 (m, 3H), 6.78 (s, 1H), 6.76 (s, 1H), 5.00-4.80 (br s, 1H), 4.80-4.60 (br s, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.50 (s, 3H), 3.48 (s, 3H), 3.20-3.05 (br s, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 161.2, 160.5, 160.4, 156.4, 156.3, 156.2, 155.5, 155.4, 154.9, 152.2, 151.9, 150.5, 150.4, 150.0, 149.1, 147.7, 144.9, 141.4, 141.1, 139.7, 138.6, 135.1, 135.0, 134.9, 134.8, 134.5, 129.9, 129.8, 129.7, 129.5, 127.0, 125.1, 125.0, 124.9, 123.9, 123.3, 122.7, 119.3, 117.8, 116.9, 116.8, 116.6, 116.3, 115.7, 114.9, 112.0, 107.9, 105.6, 61.0, 56.4, 56.0, 55.8, 41.9, 22.3.

MS (ESI) m/z: 1048 (M+1)⁺.

Rf: 0.50 (CH₂Cl₂:MeOH, 50:1).

General procedure B (starting from 153) to afford 150 as a white solid (14 mg, 88%).

¹H NMR (300 MHz, CD₃OD) δ 7.47-7.41 (m, 2H), 7.29-7.24 (m, 2H), 7.16 (s, 1H), 6.87 (d, J=8.2 Hz, 1H), 6.80 (d, J=10.1 Hz, 1H), 4.42-4.29 (m, 3H), 3.92-3.87 (m, 2H), 3.85 (s, 3H), 3.45 (s, 3H), 3.37 (s, 3H), 3.18 (t, J=6.2 Hz, 2H), 2.14-1.61 (m, 9H), 1.12-0.98 (m, 18H).

MS (ESI) m/z: 841 (M+1)⁺.

General procedure D (starting from 109 and 9H-fluorene-4-carboxylic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 200:1) to afford 151 as a white solid (26 mg, 48%).

¹H NMR (300 MHz, CDCl₃) δ 8.56-8.52 (m, 2H), 8.46-8.43 (m, 1H), 8.18-8.11 (m, 3H), 7.77-7.74 (m, 3H), 7.58-7.56 (m, 3H), 7.50-7.30 (m, 13H), 7.21 (s, 1H), 7.04 (s, 1H), 6.93 (s, 1H), 5.04-4.95 (m, 1H), 4.92-4.83 (m, 1H), 3.96 (s, 6H), 3.93 (s, 3H), 3.62 (s, 3H), 3.54 (s, 3H), 3.25 (t, J=6.5 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 166.1, 166.0, 166.0, 155.2, 152.6, 150.2, 148.0, 145.2, 145.2, 145.1, 144.2, 144.2, 144.2, 141.5, 141.3, 140.4, 139.9, 139.8, 139.2, 135.2, 134.3, 129.5, 129.3, 129.1, 129.0, 127.7, 127.3, 126.9, 126.8, 126.1, 126.1, 126.0, 125.9, 125.4, 125.1, 125.1, 125.0, 124.7, 124.6, 124.1, 123.4, 122.8, 116.3, 116.1, 115.1, 114.8, 112.2, 109.9, 105.7, 56.3, 55.9, 55.7, 42.6, 37.0 (3C), 28.2.

MS (ESI) m/z: 1100 (M+23)⁺.

Rf: 0.47 (CH₂Cl₂).

General procedure D (starting from 109 and fluorphenylsulfanylacetic acid) and chromatography on silica gel (hexane:EtOAc, 60:40) to give 152 as a white solid (47 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 7.57-7.49 (m, 6H), 7.16-7.00 (m, 10H), 6.86 (s, 1H), 6.74 (s, 1H), 6.64 (s, 1H), 4.89-4.85 (m, 1H), 4.75-4.70 (m, 1H), 3.85 (s, 2H), 3.79 (s, 4H), 3.75 (s, 3H), 3.34 (s, 3H), 3.28 (s, 3H), 3.09 (t, J=6.6 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 167.7, 167.5, 167.4, 164.2, 160.9, 154.9, 152.1, 149.7, 147.5, 144.8, 139.7, 139.2, 138.6, 134.9, 134.3, 133.8, 133.7, 133.6, 133.6, 129.3, 129.3, 126.9, 125.9, 125.8, 123.6, 123.1, 122.3, 116.4 (6C), 116.2, 116.1 (6C), 115.8, 115.0, 114.7, 111.7, 109.7, 105.5, 56.1, 55.6, 55.4, 42.4, 37.5 (3C), 27.8.

MS (ESI) m/z: 1006 (M+1)⁺.

Rf: 0.40 (hexane:EtOAc, 60:40).

General procedure D (starting from 109 and Boc-L-Leu-OH.H₂O) and chromatography on silica gel (hexane:EtOAc, 2:1) to give 153 as a white solid (77 mg, 68%).

¹H NMR (300 MHz, CDCl₃) δ 7.24-7.08 (m, 4H), 6.99 (s, 1H), 6.76 (d, J=7.0 Hz, 1H), 6.69 (d, J=8.4 Hz, 1H), 4.94-4.86 (m, 3H), 4.78-4.68 (m, 1H), 4.63-4.50 (m, 2H), 4.37-4.26 (m, 2H), 3.78 (s, 3H), 3.41 (s, 3H), 3.34 (s, 3H), 3.12 (br t, 2H), 1.90-1.60 (m, 9H), 1.45 (s, 27H), 1.05-0.95 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 177.6 (2C), 171.4, 155.6, 155.4, 155.1, 152.1, 149.7, 147.6, 144.8, 139.8, 139.2, 138.7, 135.0, 134.1, 127.0, 127.0, 126.0, 125.7, 123.8, 123.1, 122.5, 116.1, 115.8, 114.9, 114.7, 111.9, 109.7, 105.5, 80.0 (3C), 56.1, 55.7, 55.5, 53.1, 52.2 (2C), 42.4, 41.5 (3C), 28.3 (9C), 28.0, 24.7 (2C), 22.9, 22.8 (4C), 21.8 (2C).

MS (ESI) m/z: 1163 (M+23)⁺, 1141 (M+1)⁺.

Rf: 0.26 (hexane:EtOAc, 2:1).

General procedure D (starting from 26 and 5-(2-phenyleth-1-ynyl)nicotinic acid) and chromatography on silica gel (CH₂Cl₂:EtOAc, 4:1) to give 154 as a pale yellow solid (31.0 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 9.34-9.28 (m, 3H), 9.00-8.97 (m, 2H), 8.63-8.57 (m, 2H), 7.60-7.55 (m, 5H), 7.47-7.31 (m, 9H), 7.28 (s, 1H), 7.15-7.12 (m, 2H), 6.96 (s, 1H), 4.02 (s, 3H), 3.85 (s, 3H), 3.60 (s, 3H), 3.54 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 162.8, 162.6, 156.0, 154.9, 152.3, 150.4, 149.9, 149.6, 147.6, 145.5, 139.9 (3C), 139.3, 135.1, 134.2, 131.8, 129.2, 129.1, 128.5 (2C), 128.2, 124.7, 124.7, 123.9 (2C), 123.1, 122.0 (2C), 120.8, 118.9, 116.3, 115.5, 113.1, 112.1, 110.9, 108.4, 107.5, 106.3, 105.1, 94.1 (2C), 84.7 (2C), 56.3, 56.0, 55.9, 55.7.

MS (ESI) m/z: 946 (M+23)⁺, 924 (M+1)⁺.

Rf: 0.48 (CH₂Cl₂:EtOAc, 4:1).

General procedure D (starting from 95 and 5-(2-phenyleth-1-ynyl)nicotinic acid) and chromatography on silica gel (CH₂Cl₂:EtOAc, 4:1) to give 155 as a pale yellow solid (37.0 mg, 98%).

¹H NMR (300 MHz, CDCl₃) δ 9.32 (br d, J=1.9 Hz, 1H), 9.28 (br d, J=1.9 Hz, 1H), 8.99-8.97 (m, 2H), 8.60 (t, J=1.9 Hz, 1H), 8.6 (t, J=1.9 Hz, 1H), 7.59-7.55 (m, 4H), 7.40-7.36 (m, 7H), 7.26-7.21 (m, 3H), 6.84 (s, 1H), 6.80 (s, 1H), 6.75 (s, 1H), 4.95-4.75 (m, 2H), 3.92 (s, 3H), 3.82 (s, 3H), 3.51 (s, 3H), 3.50 (s, 3H), 3.16 (t, J=6.6 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 162.7, 156.0, 155.0, 152.1, 149.8, 149.2, 147.7, 147.6, 144.9, 139.9, 139.8, 139.6, 138.4, 135.9, 134.8, 131.7, 129.2, 129.1, 128.5, 128.5, 127.4, 126.5, 124.8 (2C), 123.7, 123.5, 122.0 (2C), 120.7, 119.6, 116.5, 115.0, 114.7, 114.5, 111.9, 111.0, 108.5, 105.6, 94.0, 93.9, 84.7 (2C), 56.2, 55.9, 55.8, 55.5, 42.5, 28.6.

MS (ESI) m/z: 926 (M+1)⁺.

Rf: 0.48 (CH₂Cl₂:EtOAc, 4:1).

General procedure D (starting from 109 and Boc-L-Ala-OH) and chromatography on silica gel (hexane:EtOAc, from 2:1 to 1:1) to give 156 as a white solid (81 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 7.25-7.09 (m, 4H), 6.97 (s, 1H), 6.75 (d, J=7.7 Hz, 1H), 6.67 (d, J=10.1 Hz, 1H), 5.12 (br s, 2H), 4.89-4.85 (m, 1H), 4.70-4.55 (m, 3H), 3.78 (s, 3H), 3.40 (s, 3H), 3.33 (s, 3H), 2.03 (br t, 2H), 1.58 (d, J=7.1 Hz, 3H), 1.52 (d, J=7.1 Hz, 6H), 1.47 (s, 9H), 1.45 (s, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4, 155.0, 152.0, 149.7, 147.4, 144.8, 139.7, 139.1, 138.6, 135.0, 134.1, 126.9, 126.8, 126.0, 125.9, 125.7, 123.7, 123.1, 122.4, 116.1, 115.8, 114.9, 114.7, 111.7, 109.6, 105.4, 79.9, 60.3, 56.1, 55.7, 55.7, 55.5, 55.4, 49.2, 42.3, 28.2, 27.9, 21.0, 18.5, 14.1.

MS (ESI) m/z: 1037 (M+23)⁺, 1015 (M+1)⁺.

Rf: 0.44 (hexane:EtOAc, 1:1).

General procedure E (starting from 161, reaction time 24 h) and chromatography on silica gel (hexane:EtOAc, 3:2) to give 157 as a yellow solid (27.1 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.16 (d, J=7.8 Hz, 1H), 7.60-7.50 (m, 6H), 7.22-7.20 (m, 1H), 7.10-7.00 (m, 10H), 6.75 (s, 2H), 3.93 (s, 2H), 3.87 (s, 2H), 3.82 (s, 3H), 3.80 (s, 2H), 3.79 (s, 3H), 3.45 (s, 3H), 3.37 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 167.8, 167.7, 167.7, 164.6, 164.5, 161.3, 161.2, 155.1, 153.4, 152.5, 147.8, 145.6, 142.1, 140.3, 139.8, 138.9, 134.8, 134.1, 134.0, 133.4, 128.5, 124.0, 123.9, 123.7, 121.2, 118.3, 116.9, 116.6, 116.5, 116.4, 116.1, 115.4, 115.4, 112.3, 112.2, 112.1, 109.2, 106.7, 106.5, 104.5, 61.2, 56.5, 55.9, 55.8, 37.8. MS (APCI) m/z: 1034 (M+1)⁺.

Rf: 0.63 (hexane:EtOAc, 3:2).

General procedure E (starting from 74, reaction time 6 d) and chromatography on silica gel (hexane:EtOAc, 2:3) to give 158 as a yellow solid (48.3 mg, 68%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.5 Hz, 1H), 7.33-7.06 (m, 21H), 6.77 (d, J=8.4 Hz, 1H), 5.09 (s, 6H), 4.94-4.91 (m, 3H), 4.56 (m, 3H), 3.85 (s, 3H), 3.79 (s, 3H), 3.48 (s, 3H), 3.41 (s, 3H), 3.25-3.24 (m, 6H), 2.19-1.82 (m, 6H), 1.62-1.50 (m, 12H), 1.46-1.45 (m, 27H).

¹³C NMR (75 MHz, CDCl₃) δ 171.5, 170.9 (2C), 156.8 (3C), 156.0, 155.7, 155.1, 153.3 (2C), 152.5, 147.8, 145.6 (2C), 141.9, 140.2, 139.6 (2C), 139.1 (2C), 136.8, 136.7, 136.2, 134.8, 133.5 (2C), 128.8, 128.3, 124.3, 123.9, 123.7, 121.2, 118.5, 116.1, 115.4, 112.3, 109.2, 107.1, 106.4, 104.6, 80.6, 80.4 (2C), 66.9 (3C), 61.1, 56.5, 56.0 (2C), 54.0, 53.7 (2C), 40.8, 40.7 (2C), 32.3, 32.0 (2C), 29.8 (3C), 28.6 (9C), 22.6, 22.5 (2C).

MS (ESI) m/z: 1638 (M+23)⁺.

Rf: 0.44 (hexane:EtOAc, 2:3).

General procedure E (starting from 76, reaction time 6 d) and chromatography on silica gel (hexane:EtOAc, 2:1) to give 159 as a yellow solid (27.9 mg, 69%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.6 Hz, 1H), 7.32-7.08 (m, 6H), 6.80-6.77 (m, 1H), 5.01-4.98 (m, 3H), 6.60 (m, 3H), 3.84 (s, 3H), 3.81 (s, 3H), 3.48 (s, 3H), 3.43 (s, 3H), 1.91-1.62 (m, 9H), 1.50-1.46 (m, 27H), 1.06-0.99 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 171.4, 155.6, 155.5, 155.3, 155.0, 153.2, 152.3, 147.6, 145.5, 145.4, 141.7, 140.1, 139.6, 139.0, 134.5, 133.2 (2C), 128.3, 124.0, 123.6, 123.5, 121.0, 118.4, 115.8, 115.1, 112.1, 109.0, 107.0, 106.2, 104.2, 80.4, 80.1 (2C), 56.2, 55.8, 55.7, 55.6, 52.6, 52.2 (2C), 41.7, 41.5, 41.3, 28.3 (9C), 24.8 (3C), 23.0, 22.9 (3C), 21.9, 21.8.

MS (ESI) m/z: 1191 (M+23)⁺.

Rf: 0.55 (hexane:EtOAc, 2:1).

General procedure E (starting from 75, reaction time 3 d) and chromatography on silica gel (hexane:EtOAc, 2:1) to give 160 as a yellow solid (37.8 mg, 77%).

¹H NMR (300 MHz, CDCl₃) δ 9.22 (d, J=7.6 Hz, 1H), 7.33-7.09 (m, 6H), 6.78 (d, J=8.8 Hz, 1H), 5.12-5.06 (m, 3H), 4.63-4.53 (m, 3H), 3.86 (s, 3H), 3.81 (s, 3H), 3.49 (s, 3H), 3.43 (s, 3H), 2.46-2.35 (m, 3H), 1.49-1.44 (m, 27H), 1.31-1.01 (m, 18H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4 (3C), 155.9, 155.7, 154.9, 153.1 (2C), 152.2, 147.6, 145.4 (2C), 141.7, 139.9, 138.7, 134.6, 133.2 (2C), 124.0, 123.6, 123.5, 121.0, 118.3, 115.8, 115.1, 112.1 (2C), 109.0, 106.9, 106.1, 104.2, 80.3, 79.9 (2C), 60.7, 59.0, 58.5 (2C), 56.0, 55.7, 55.6, 31.3, 31.1, 30.9, 28.3 (9C), 19.3, 19.2, 19.0, 17.5, 17.2, 17.1.

MS (ESI) m/z: 1149 (M+23)⁺, 1127 (M+1)⁺.

Rf: 0.42 (hexane:EtOAc, 2:1).

General procedure D (starting from 1 and 2-[(4-fluorophenyl)thio]acetic acid) and chromatography on silica gel (hexane:EtOAc, 3:2) to give a yellow solid which contained 2-[(4-fluorophenyl)thio]acetic acid. The solid was dissolved in CH₂Cl₂ (20 mL) and washed with NaOH 1 M (20 mL) to give 161 as a pale yellow solid (52.3 mg, 54%).

¹H NMR (300 MHz, CDCl₃) δ 7.57-7.49 (m, 6H), 7.16-7.00 (m, 10H), 6.64 (s, 1H), 6.63 (s, 1H), 4.80-4.76 (m, 1H), 4.70-4.55 (m, 1H), 3.87 (s, 2H), 3.85 (s, 2H), 3.79 (s, 2H), 3.77 (s, 3H), 3.74 (s, 3H), 3.35 (s, 3H), 3.34 (s, 3H), 2.90 (br t, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 167.6, 167.5, 167.4, 164.2, 160.9, 154.8, 152.1, 151.8, 147.5, 144.8, 141.3, 141.1, 139.7, 138.6, 134.7, 134.3, 133.7 (2C), 133.6, 133.5, 133.4, 129.3 (2C), 126.9, 123.5, 123.2, 122.6, 119.0, 116.5, 116.3, 116.2, 116.1, 116.0, 115.6, 114.7, 111.6, 107.6, 105.5, 60.8, 56.2, 55.6, 55.4, 41.8, 37.5, 37.4, 37.3, 29.6.

MS (ESI) m/z: 1057 (M+23)⁺, 1035 (M+1)⁺.

Rf: 0.71 (hexane:EtOAc, 1:1).

General procedure G (starting from 6,7-Dimethoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (hexane:EtOAc, 2:1) to afford 162 as a pale yellow solid (274.8 mg, 47%).

¹H NMR (300 MHz, CDCl₃) δ 7.11-7.03 (m, 3H), 6.91 (s, 1H), 6.75 (s, 1H), 6.73 (s, 1H), 6.67 (s, 1H), 4.83-4.61 (m, 2H), 4.59-4.51 (m, 2H), 3.89 (s, 3H), 3.82 (s, 3H), 3.42, 3.36 (s, 3H), 3.12 (t, J=6.8 Hz, 2H), 1.39-1.36 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.2, 152.0, 148.6, 147.1, 146.6, 146.1, 145.5, 135.5, 128.2, 127.8, 126.3, 123.1, 119.7, 116.6, 114.5, 114.4, 113.3, 110.7, 110.0, 108.3, 104.5, 103.0, 71.4, 71.0, 55.9, 55.6, 55.1, 54.7, 42.0, 28.3, 21.6, 21.5, 21.5, 21.4.

MS (ESI) m/z: 600 (M+1)⁺.

Rf: 0.17 (hexane:EtOAc, 2:1).

General procedure G (starting from 6-Benzyloxy-7-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (hexane:EtOAc, 2:1) to afford 163 as a pale yellow solid (42.5 mg, 34%).

¹H NMR (300 MHz, CDCl₃) δ 7.43-7.29 (m, 5H), 7.09-7.03 (m, 3H), 6.90 (s, 1H), 6.76 (s, 1H), 6.75 (s, 1H), 6.66 (s, 1H), 5.14 (s, 2H), 4.79-4.50 (m, 4H), 3.82 (s, 3H), 3.42 (s, 3H), 3.37 (s, 3H), 3.04 (t, J=6.8 Hz, 2H), 1.39-1.36 (m, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 151.2, 148.0, 148.0, 147.0, 146.9, 146.5, 145.9, 136.6, 135.8, 128.6, 128.5, 128.2, 128.0, 127.1, 126.4, 125.4, 123.4, 120.5, 116.9, 114.9, 114.5, 113.7, 113.3, 110.3, 109.0, 108.5, 104.8, 103.4, 71.7, 71.4, 70.9, 56.1, 55.4, 55.1, 42.3, 28.6, 21.8 (4C).

MS (ESI) m/z: 676 (M)⁺.

Rf: 0.30 (hexane:EtOAc, 2:1).

General procedure L (starting from 95) to afford 164 as a brown solid (7 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 7.23-7.09 (m, 4H), 6.76 (s, 1H), 6.72 (s, 1H), 6.69 (s, 1H), 4.90-4.74 (m, 2H), 4.30 (t, J=6.6 Hz, 2H), 3.90 (s, 3H), 3.80 (s, 3H), 3.43 (s, 3H), 3.16 (s, 3H), 3.14 (t, J=6.6 Hz, 2H), 2.34 (s, 3H), 2.31 (s, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 168.7, 152.1, 149.1, 147.7 (2C), 139.9, 138.8, 134.3, 130.9, 128.8, 126.4, 126.3, 123.8, 123.3, 119.7, 116.2, 114.8 (2C), 111.9, 111.0, 108.5, 105.5, 56.2, 55.9, 55.7, 55.4, 42.5, 29.7 (2C), 29.4.

MS (ESI) m/z: 600 (M+1)⁺.

Rf: 0.27 (EtOAc:hexane, 2:1).

General procedure G (starting from 7-Isopropoxy-6-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (hexane:EtOAc, 50:50) to afford 165 as a pale yellow solid (84.6 mg, 23%).

¹H NMR (300 MHz, CDCl₃) δ 7.08-7.00 (m, 3H), 6.86 (s, 1H), 6.74 (s, 2H), 6.60 (s, 1H), 4.78-4.71 (m, 2H), 4.60-4.41 (m, 2H), 3.88-3.77 (m, 1H), 3.83 (s, 3H), 3.80 (s, 3H), 3.40 (s, 3H), 3.08 (t, J=6.8 Hz, 2H), 1.40 (d, J=6.1 Hz, 6H), 1.33 (d, J=6.0 Hz, 6H), 1.10 (d, J=6.1 Hz, 3H), 1.07 (d, J=6.1 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 155.5, 151.1, 149.9, 147.0, 146.8, 146.4, 145.8, 145.7, 135.8, 128.3, 128.2, 126.4, 123.2, 119.9, 116.2, 114.7, 114.3, 113.5, 111.9, 111.4, 110.3, 104.8, 103.3, 71.5, 71.3, 70.7, 55.9, 55.8, 55.3, 42.3, 28.6, 21.9, 21.8, 21.7, 21.5(2C), 20.9.

MS (ESI) m/z: 650 (M+23)⁺, 628 (M+1)⁺.

Rf: 0.41 (hexane:EtOAc, 50:50).

General procedure L (starting from 26) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 166 (7 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 9.30 (d, J=7.6 Hz, 1H), 7.31-7.26 (m, 5H), 7.23-7.10 (m, 2H), 6.84 (s, 1H), 4.00 (s, 3H), 3.83 (s, 3H), 3.51 (s, 3H), 3.46 (s, 3H), 2.37 (s, 2H), 2.32 (s, 2H).

MS (ESI) m/z: 620 (M+23)⁺, 598 (M+1)⁺.

Rf: 0.60 (CH₂Cl₂:MeOH, 10:1).

General procedure A (starting from 165) and chromatography on silica gel (CH₂Cl₂:MeOH, 20:1) to afford 167 as a beige solid (35.3 mg, 55%).

¹H NMR (300 MHz, DMSO-d₆) δ 9.66 (br s, 1H), 9.26 (br s, 1H), 8.85 (br s, 1H), 6.99-6.94 (m, 3H), 6.83 (dd, J=7.8, 1.7 Hz, 1H), 6.78 (s, 1H), 6.66 (s 1H), 6.46 (s, 1H), 4.62 (br t, J=5.9 Hz, 2H), 3.79 (s, 3H), 3.73 (s, 3H), 3.33 (s, 3H), 3.06 (br t, J=5.9 Hz, 2H).

¹³C NMR (75 MHz, DMSO-d₆) δ 154.3, 148.4, 148.0, 146.8, 146.5, 145.6, 144.8, 144.4, 135.4, 127.9, 125.5, 125.3, 123.3, 119.7, 116.4, 114.8, 114.5, 112.8, 112.4, 111.9, 108.8, 105.0, 103.5, 55.9, 55.6, 55.0, 42.0, 28.0.

MS (ESI) m/z: 524 (M+23)⁺, 502 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂:MeOH, 20:1).

General procedure D (starting from 3 and 6-(BOC-amino)caproic acid) and chromatography on silica gel (CH₂Cl₂:MeOH, 40:1) to afford 168 as a white solid (608 mg, 89%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.3 Hz, 1H), 7.38 (s, 1H), 7.29-7.13 (m, 5H), 7.07 (d, J=7.5 Hz, 1H), 6.80 (s, 1H), 4.56 (bs, 3H), 3.82 (s, 3H), 3.44 (s, 6H), 3.20-3.13 (m, 6H), 2.66-2.56 (m, 6H), 1.84-1.75 (m, 6H), 1.60-1.44 (m, 39H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4, 171.2, 155.9, 155.0, 152.4, 151.0, 147.7, 145.4, 140.9, 140.3, 139.8, 134.1, 133.5, 128.2, 124.0, 123.8, 123.6, 123.5, 123.0, 120.6, 115.5, 115.0, 112.7, 112.2, 112.1, 108.9, 106.3, 106.1, 79.0(3C), 56.2, 55.7, 55.6, 40.3(3C), 33.8(2C), 33.7, 29.7(3C), 28.4(9C), 26.2, 26.1(2C), 24.5(2C), 24.5.

MS (ESI) m/z: 1162 (M+23)⁺.

Rf: 0.30 (CH₂Cl₂:MeOH, 40:1).

General procedure C (starting from 168) to afford 169 as a white solid (389 mg, 93%).

¹H NMR (300 MHz, CD₃OD) δ 9.07 (d, J=7.3 Hz, 1H), 7.49 (s, 2H), 7.39 (d, J=8.1 Hz, 1H), 7.31-7.28 (m, 2H), 7.16-7.11 (m, 2H), 6.86 (s, 1H), 3.88 (s, 3H), 3.46 (s, 6H), 3.02-2.94 (m, 6H), 2.73-2.60 (m, 6H), 1.88-1.75 (m, 12H), 1.54-1.52 (m, 6H).

MS (ESI) m/z: 839 (M+1)⁺.

Me₃SiCl (12 mL, 0.095 mmol) was added to a suspension of 57 (7.0 mg, 0.0136 mmol) in MeOH (2 mL). The solution was stirred at 23° C. for 1 hour. The solvent was evaporated to dryness and CH₂Cl₂ (2×1 mL) was added in order to remove all the solvent to give 170 as a light orange solid (8 mg, quant.).

¹H NMR (300 MHz, CD₃OD) δ 7.70 (d, J=7.3 Hz, 1H), 7.47 (s, 1H), 7.42 (s, 1H), 7.32 (d, J=7.3 Hz, 1H), 6.96 (s, 1H), 6.79 (s, 1H), 6.56 (s, 1H), 4.90-4.79 (m, 4H), 3.98 (s, 3H), 3.85 (s, 3H), 3.57 (s 3H), 3.34 (s, 3H).

General procedure G (starting from 4-bromoisoquinoline) and chromatography on silica gel (hexane:CH₂Cl₂:Et₂O, 6:4:1) to provide 171 as a yellow solid (41 mg, 16%).

¹H NMR (300 MHz, CDCl₃) δ 9.61 (s, 1H), 8.11 (d, J=7.8 Hz, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.59 (t, J=7.7 Hz, 1H), 7.34 (t, J=7.8 Hz, 1H), 7.17 (d, J=8.0 Hz, 1H), 7.12-7.05 (m, 2H), 6.96 (s, 1H), 6.64 (s, 1H), 4.70 (hp, J=6.0 Hz, 1H), 4.57 (hp, J=6.0 Hz, 1H), 3.83 (s, 3H), 3.44 (s, 3H), 1.51 (d, J=6.2 Hz, 3H), 1.44 (d, J=6.0 Hz, 3H), 1.40 (d, J=7.8 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 151.5, 148.1, 147.5, 146.7, 146.6, 133.3, 129.8, 128.8, 128.4, 127.6, 127.0, 125.5, 125.0, 124.6, 123.2, 116.6, 114.8, 114.2, 113.5, 109.5, 108.8, 108.4, 105.4, 103.2, 71.5, 56.1, 55.4, 22.3, 21.9, 21.8, 21.7.

Rf: 0.46 (hexane:CH₂Cl₂:Et₂O, 6:4:1).

General procedure A (starting from 171) and chromatography on silica gel (CH₂Cl₂:MeOH from 30:1 to 10:1) to afford 172 as a yellow solid (20 mg, 74%).

¹H NMR (300 MHz, (CD₃)₂SO, 40° C.) δ 9.85 (s, 1H), 9.38 (s, 1H), 9.36 (s, 1H), 8.03 (d, J=8.4 Hz, 1H), 7.79 (d, J=8.2 Hz, 1H), 7.70 (t, J=7.3 Hz, 1H), 7.54 (t, J=7.3 Hz, 1H), 7.20-7.05 (m, 2H), 6.96 (d, J=7.8 Hz, 1H), 6.87 (s, 1H), 6.55 (s, 1H), 3.74 (s, 3H), 3.37 (s, 3H).

¹³C NMR (75 MHz, (CD₃)₂SO, 60° C.) δ 154.1, 148.8, 148.1, 147.1, 146.2, 144.7, 132.2, 129.4, 129.1, 128.6, 127.2, 126.2, 124.4, 123.9, 123.0, 116.7, 114.5, 113.5, 107.7, 107.3, 105.8, 103.6, 72.0, 55.9, 55.0. MS (APCI) m/z: 534 (M+2)⁺, 532 (M)⁺.

Rf: 0.50 (CH₂Cl₂:MeOH, 20:1).

General procedure D (starting from 3 and Boc-Lys(Boc)Gly-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 30:1) to afford 173 as a pale yellow solid (98 mg, 74%).

¹H NMR (300 MHz, CDCl₃) δ 8.62 (d, J=7.3 Hz, 1H), 7.70-7.40 (m, 3H), 7.30-7.00 (m, 6H), 6.80-6.60 (m, 2H), 6.40 (d, J=7.1 Hz, 1H), 5.67 (br s, 2H), 5.41 (br s, 1H), 4.91 (br s, 1H), 4.82 (br s, 2H), 4.50-4.20 (m, 9H), 3.90 (s, 3H), 3.42 (s, 6H), 3.05 (br s, 6H), 1.90-1.60 (m, 6H), 1.50-1.30 (m, 66H).

¹³C NMR (75 MHz, CDCl₃) δ 173.2, 172.8, 167.8, 167.7, 156.3, 156.1, 155.9, 154.1, 152.1, 150.6, 147.5, 144.7, 140.0, 139.6, 138.9, 134.4, 132.9, 127.4, 123.7, 123.4, 123.3, 121.9, 121.0, 115.4, 112.2, 111.9, 108.2, 106.0, 105.7, 79.9, 78.9, 56.4, 55.7, 55.5, 54.3, 53.4, 40.9, 39.8, 31.9, 29.6, 28.4, 28.3, 28.2, 27.7, 22.5. MS (APCI) m/z: 1678 (M+23)⁺.

Rf: 0.21 (CH₂Cl₂:MeOH, 30:1).

General procedure H (starting from 6,7-dimethoxy-5-isopropoxy-3,4-dihydroisoquinoline) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 100:1) to afford 174 as a clear oil (150 mg, 43%).

¹H NMR (300 MHz, CDCl₃) δ 7.16 (s, 1H), 7.00 (s, 1H), 6.92 (s, 1H), 6.81 (s, 1H), 4.70-4.50 (m, 4H), 3.95 (s, 6H), 3.88 (s, 3H), 3.10 (t, J=6.7 Hz, 2H), 1.41 (d, J=6.2 Hz, 6H), 1.31 (d, J=6.2 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 152.7, 148.9, 147.6, 147.2, 145.9, 143.2, 139.7, 130.9, 122.8, 120.4, 115.1, 109.9, 104.6, 103.6, 103.4, 96.0, 75.6, 71.5, 60.6, 56.4, 56.1, 42.0, 22.6, 22.5, 21.8.

MS (ESI) m/z: 494 (M+1)⁺.

Rf: 0.40 (CH₂Cl₂:MeOH, 100:1).

Gerenal procedure A (starting from 174) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 50:1) to afford 175 as a brown solid (15 mg, 62%).

¹H NMR (300 MHz, CDCl₃) δ 7.16 (s, 1H), 6.99 (s, 1H), 6.84 (s, 1H), 6.81 (s, 1H), 5.99 (s, 1H), 5.84 (s, 1H), 4.70 (t, J=6.9 Hz, 2H), 4.01 (s, 3H), 3.96 (s, 6H), 3.11 (t, J=6.9 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 151.3, 146.6, 146.4, 146.0, 143.9, 139.6, 136.1, 130.9, 123.1, 115.3, 112.4, 109.9, 103.7, 103.4, 100.1, 96.0, 61.2, 56.4, 56.0, 41.9, 21.3.

MS (ESI) m/z: 410 (M+1)⁺.

Rf: 0.44 (CH₂Cl₂:MeOH, 20:1).

Gerenal procedure H (starting from 5-Boc-aminoisoquinoline) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH:Et₃N, 100:1:0.5) to afford 176 as a brown solid (120 mg, 10%).

¹H NMR (300 MHz, CDCl₃) δ 9.61 (s, 1H), 8.11 (d, J=7.8 Hz, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.59 (t, J=7.7 Hz, 1H), 7.34 (t, J=7.8 Hz, 1H), 7.17 (d, J=8.0 Hz, 1H), 7.12-7.05 (m, 2H), 6.96 (s, 1H), 6.64 (s, 1H), 4.70 (hp, J=6.0 Hz, 1H), 4.57 (hp, J=6.0 Hz, 1H), 3.83 (s, 3H), 3.44 (s, 3H), 1.51 (d, J=6.2 Hz, 3H), 1.44 (d, J=6.0 Hz, 3H), 1.40 (d, J=7.8 Hz, 6H).

MS (ESI) m/z: 653 (M+1)⁺.

Rf: 0.33 (CH₂Cl₂:MeOH, 100:1).

General procedure I (starting from 3) and chromatography on silica gel (CH₂Cl₂) and triturated with Et₂O (50 mL) to afford 177 as a white solid (434.7 mg, 82%).

¹H NMR (300 MHz, CDCl₃) δ 9.31 (d, J=7.3 Hz, 1H), 7.62 (s, 1H), 7.57 (d, J=8.5 Hz, 1H), 7.35-7.32 (m, 3H), 7.16 (d, J=7.6 Hz, 1H), 7.14 (s, 1H), 6.74 (s, 1H), 3.97 (s, 3H), 3.50 (s, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 154.3, 152.9, 150.9, 148.1, 145.0, 139.4, 139.0, 138.1, 136.7, 132.7, 127.3, 124.9, 124.0, 123.8, 123.7, 121.0, 120.8, 117.5, 116.5, 115.8, 113.2, 112.3, 109.9, 106.8, 106.4, 56.8, 55.9, 55.7.

MS (ESI) m/z: 896 (M+1)⁺.

Rf: 0.32 (hexane:EtOAc, 6:1).

AlCl₃ (12 mg, 0.092 mmol) was added to a solution of 176 (20 mg, 0.030 mmol) in anhydrous CH₂Cl₂ (2 mL) under Argon atmosphere. The reaction mixture was stirred for 2.5 hours at 23° C. The mixture was quenched with H₂O (10 mL, pH=4-5), extracted with CH₂Cl₂ (3×10 mL), dried over anhydrous Na₂SO₄ and evaporated under reduced pressure.

The resulted residue was subjected to flash chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 30:1) to provide 178 as a white solid (7 mg, 42%).

¹H NMR (300 MHz, CDCl₃) δ 9.28 (d, J=7.7 Hz, 1H), 7.25-7.05 (m, 6H), 6.97 (s, 1H), 6.82 (d, J=7.7 Hz, 1H), 6.65 (s, 1H), 4.69 (hp, J=6.2 Hz, 1H), 4.57 (hp, J=6.2 Hz, 1H), 4.12 (bs, 2H), 3.82 (s, 3H), 3.43 (s, 3H), 1.49 (d, J=5.9 Hz, 3H), 1.43 (d, J=5.9 Hz, 3H), 1.40 (d, J=6.2 Hz, 6H).

MS (ESI) m/z: 553 (M+1)⁺.

Rf: 0.55 (CH₂Cl₂:MeOH, 30:1).

General procedure H (starting from 7-isopropylisoquinoline) and chromatography on reverse silica gel RP-18 (CH₃CN:H₂O, 4:1 then CH₃CN) to afford 179 as a yellow oil (6 mg, 2%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.3 Hz, 1H), 7.61 (d, J=7.3 Hz, 1H), 7.17-7.05 (m, 6H), 6.98 (s, 1H), 6.70 (s, 1H), 4.65-4.58 (m, 2H), 4.1-3.95 (m, 1H), 3.83 (s, 3H), 3.44 (s, 3H), 1.47 (d, J=6.1 Hz, 6H), 1.40 (d, J=6.1 Hz, 6H), 1.17-1.12 (m, 6H).

MS (ESI) m/z: 596 (M+1)⁺.

Rf: 0.31 (CH₃CN, RP-18).

General procedure C (starting from 127) to afford 180 as a pale yellow solid (156 mg, 88%).

¹H NMR (300 MHz, CD₃OD) δ 9.17 (dd, J=7.6, 2.6 Hz, 1H), 7.69 (d, J=2.7 Hz, 1H), 7.65-7.55 (m, 2H), 7.50-7.20 (m, 4H), 6.90 (d, J=10.6 Hz, 1H), 4.80-4.60 (m, 3H), 3.92 (s, 3H), 3.91 (s, 3H), 3.60-3.40 (m, 12H), 2.70-2.30 (m, 6H), 2.30-10 (m, 6H).

MS (ESI) m/z: 791 (M+1)⁺.

General procedure C (starting from 146) to afford 181 as a white solid (390 mg, 84%).

¹H NMR (300 MHz, CD₃OD) δ 9.26 (d, J=7.5 Hz, 1H), 7.69 (s, 1H), 7.60-7.50 (m, 2H), 7.45-7.30 (m, 4H), 6.92 (d, J=6.6 Hz, 1H), 4.50-4.30 (m, 3H), 3.88 (s, 3H), 3.48 (s, 6H), 2.20-1.70 (m, 6H), 1.20-1.00 (m, 18H).

MS (ESI) m/z: 840 (M+1)⁺.

General procedure A (starting from 178) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 30:1) to afford 182 as a white solid (3.8 mg, 76%).

¹H NMR (300 MHz, CD₃OD) δ 9.15 (d, J=7.7 Hz, 1H), 7.20-7.15 (m, 2H), 7.10-6.95 (m, 4H), 6.89 (s, 1H), 6.82 (d, J=7.7 Hz, 1H), 6.57 (s, 1H), 3.81 (s, 3H), 3.44 (s, 3H).

MS (ESI) m/z: 469 (M+1)⁺.

Rf: 0.12 (CH₂Cl₂:MeOH, 40:1).

A suspension of 177 (0.33 g, 0.37 mmol), Pd(OAc)₂ (12.5 mg, 0.055 mmol), BINAP (69.2 mg, 0.111 mmol) in anhydrous toluene (5 mL) was stirred at 23° C. under Argon atmosphere for 5 min. Then benzophenone imine (218 mL, 1.30 mmol) was added and the mixture was stirred at 110° C. for 7 days. The reaction was cool down to 23° C., H₂O (20 mL) was added, was extracted with CH₂Cl₂ (3×20 mL), dried over anhydrous Na₂SO₄, filtered and evaporated to dryness. The residue was purified by chromatography on silica gel (hexane:EtOAc, 2:1) to give 183 (29.0 mg, 8%) as a yellow solid.

¹H NMR (300 MHz, CDCl₃) δ 9.11 (d, J=7.5 Hz, 1H), 7.78-7.70 (m, 4H), 7.48-7.13 (m, 26H), 7.07-6.97 (m, 3H), 6.86-6.80 (m, 3H), 6.67 (s, 1H), 6.64 (s, 1H), 3.69 (s, 3H), 3.28 (s, 3H), 3.26 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 170.4, 169.3, 155.5, 150.3, 150.0, 146.4, 146.0, 142.8, 141.9, 140.9, 139.4, 139.0, 137.0, 136.4, 134.2, 131.1, 130.9, 129.6, 129.4, 128.8, 128.6, 128.5, 128.2, 127.9, 127.9 (4C), 124.1, 123.8, 122.7, 121.7, 121.1, 117.5, 114.4, 113.1, 112.6, 111.9, 109.2, 105.4, 105.2, 55.7, 55.6, 55.3.

MS (ESI) m/z: 989 (M+1)⁺.

Rf: 0.50 (Hex:EtOAc, 2:1).

General procedure G (starting from 7-hydroxy-isoquinoline) and chromatography on silica gel (CH₂Cl₂:EtOAc, 200:1) to afford 184 as a white solid (112.5 mg, 9%).

¹H NMR (300 MHz, (CD₃)₂SO) δ 10.0 (br s, 1H), 8.84 (d, J=7.0 Hz, 1H), 7.61 (d, J=8.8 Hz, 1H), 7.09-7.01 (m, 3H), 6.71 (s, 2H), 6.6 (s, 1H), 6.5 (s, 1H), 4.64 (m, 1H), 4.42 (m, 1H), 3.33 (s, 3H), 3.31 (s, 3H), 1.27 (d, J=5.7 Hz, 6H), 1.13 (d, J=5.7 Hz, 6H).

¹³C NMR (75 MHz, (CD₃)₂SO) δ 158.9, 157.0, 155.4, 149.2, 147.6, 147.2, 147.1, 145.7, 133.8, 128.6, 124.5, 121.5, 121.3, 118.6, 114.6, 114.5, 113.9, 110.8, 110.0, 108.3, 106.9, 101.5, 92.8, 83.8, 70.5, 70.1, 55.9, 54.9, 54.8, 21.7 (2C).

MS (ESI) m/z: 575 (M+1)⁺.

Rf: 0.23 (CH₂Cl₂:EtOAc, 200:1).

General procedure H (starting from 3,4-dihydroisoquinoline) and chromatography on silica gel (CH₂Cl₂ and then hexane:EtOAc, 2:1) to afford 185 as a pale yellow solid (243 mg, 21%).

¹H NMR (300 MHz, CDCl₃) δ 7.28-7.00 (m, 7H), 6.91 (s, 1H), 6.63 (s, 1H), 4.80-4.78 (m, 2H), 4.64 (sep, J=6.0 Hz, 1H), 4.53 (sep, J=6.0 Hz, 1H), 3.81 (s, 3H), 3.42 (s, 3H), 317 (t, J=6.5 Hz, 2H), 1.39-1.37 (m, 12H).

MS (ESI) m/z: 541 (M+1)⁺.

Rf: 0.50 (hexane:EtOAc, 2:1).

General procedure H (starting from 6-isopropoxy-7-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, from 100:1 to 20:1) to afford 186 as a brown solid (861 mg, 29%).

¹H NMR (300 MHz, CDCl₃) δ 7.17 (s, 1H), 7.15 (s, 1H), 6.90 (s, 1H), 6.78 (s, 1H), 6.77 (s, 1H), 4.68 (t, J=6.7 Hz, 2H), 4.62-4.50 (m, 2H), 3.94 (s, 3H), 3.93 (s, 3H), 3.06 (t, J=6.7 Hz, 2H), 1.40 (d, J=6.0 Hz, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 149.6, 148.1, 147.6, 147.2, 145.9, 140.1, 131.1, 125.6, 120.0, 115.0, 114.9, 110.0, 108.1, 104.6, 103.6, 95.3, 71.5, 56.4, 56.2, 42.2, 28.2, 22.0, 21.8.

MS (ESI) m/z: 464 (M+1)⁺.

Rf: 0.44 (hexane:AcOEt, 1:1).

N-Bromosuccinimide (21 mg, 0.12 mmol) was added in one portion to a solution of 186 (50 mg, 0.10 mmol) in AcOEt (1 mL) under Argon atmosphere. The solution was stirred at 23° C. for 15 minutes, then diluted with AcOEt, quenched with H₂O and washed succesively with HCl 0.1N (2×10 mL) and NaOH 0.1N (2×10 mL).

After drying over Na₂SO₄, the solvent was evaporated under vacuum to afford 187 as a brown solid (56 mg, 96%).

¹H NMR (300 MHz, CDCl₃) δ 8.19 (s, 1H), 8.12 (s, 1H), 6.89 (s, 1H), 6.81 (s, 1H), 4.73 (t, J=6.2 Hz, 2H), 4.65-4.50 (m, 2H), 3.94 (s, 3H), 3.93 (s, 3H), 3.02 (t, J=6.5 Hz, 2H), 1.41 (d, J=6.0 Hz, 6H), 1.40 (d, J=6.0 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 154.7, 148.8, 148.0, 147.6, 146.5, 145.9, 135.2, 127.3, 127.0, 119.2, 114.6, 114.0, 109.6, 109.5, 104.7, 103.2, 86.5, 71.4, 56.3, 56.2, 42.5, 28.8, 22.0, 21.8.

MS (ESI) m/z: 564 (M+23)⁺.

Rf: 0.58 (hexane:AcOEt, 1:1).

General procedure A (starting from 187) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, from 100:1 to 40:1) to afford 188 as a brown solid (15 mg, 40%).

¹H NMR (300 MHz, CDCl₃) δ 8.23 (s, 1H), 8.11 (s, 1H), 6.87 (s, 1H), 6.79 (s, 1H), 4.67 (t, J=6.6 Hz, 2H), 3.96 (s, 3H), 3.94 (s, 3H), 2.97 (t, J=6.6 Hz, 2H).

MS (ESI) m/z: 458 (M+1)⁺.

Rf: 0.14 (CH₂Cl₂:MeOH, 50:1).

N-Iodosuccinimide (77 mg, 0.32 mmol) was added in one portion to a solution of 186 (100 mg, 0.21 mmol) in CH₂Cl₂ (4 mL) under Argon atmosphere. The solution was stirred at 23° C. for 30 minutes, then diluted with AcOEt, quenched with H₂O and washed succesively with NaOH 0.1N (2×10 mL) and H₂O (2×10 mL).

After drying over Na₂SO₄, the solvent was evaporated under vacuum to afford 189 as a brown solid (120 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 8.48 (s, 1H), 8.25 (s, 1H), 6.87 (s, 1H), 6.81 (s, 1H), 4.74 (t, J=6.2 Hz, 2H), 4.65-4.50 (m, 2H), 3.97 (s, 3H), 3.94 (s, 3H), 2.99 (t, J=6.4 Hz, 2H), 1.41 (d, J=6.0 Hz, 6H), 1.40 (d, J=6.0 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 154.5, 148.4, 148.0, 147.5, 145.9, 137.5, 129.4, 127.7, 119.5, 115.7, 114.6, 110.0, 103.7, 103.2, 71.3, 56.3, 42.5, 29.0, 22.0, 21.8.

MS (ESI) m/z: 590 (M+1)⁺.

Rf: 0.49 (hexane:AcOEt, 1:1).

General procedure C (starting from 129) to afford 190 as a white solid (197 mg, 80%).

¹H NMR (300 MHz, CD₃OD) δ 9.23 (d, J=7.5 Hz, 1H), 7.60-7.50 (m, 2H), 7.45-7.30 (m, 3H), 7.24 (d, J=7.4 Hz, 1H), 6.92 (d, J=10.4 Hz, 1H), 4.60 (d, J=3.7 Hz, 1H), 4.36 (d, J=4.3 Hz, 1H), 4.27 (d, J=4.3 Hz, 1H), 3.90 (d, J=1.2 Hz, 3H), 3.89 (d, J=2.4 Hz, 3H), 3.54 (d, J=3.8 Hz, 3H), 3.48 (d, J=3.5 Hz, 3H), 2.70-2.40 (m, 3H), 1.35-1.15 (m, 18H).

MS (ESI) m/z: 827 (M+1)⁺.

General procedure C (starting from 97) to afford 191 as a white solid (1.15 g, 94%).

¹H NMR (300 MHz, CD₃OD) δ 9.16 (d, J=7.7 Hz, 1H), 7.60-7.50 (m, 2H), 7.40-7.25 (m, 3H), 7.22 (d, J=7.4 Hz, 1H), 6.88 (d, J=9.1 Hz, 1H), 4.70-4.60 (m, 1H), 4.60-4.50 (m, 1H), 4.50-4.35 (m, 1H), 3.91 (s, 3H), 3.90 (s, 3H), 3.54 (d, J=2.1 Hz, 3H), 3.48 (d, J=2.1 Hz, 3H), 1.90-1.70 (m, 9H).

MS (ESI) m/z: 743 (M+1)⁺.

General procedure D (starting from 2 and 6-(BOC-amino)caproic acid) and chromatography on silica gel (hexane:EtOAc, 50:50) to afford 192 as a white solid (2.02 g, 92%).

¹H NMR (300 MHz, CDCl₃) δ 9.17 (d, J=7.5 Hz, 1H), 7.30-7.20 (m, 3H), 7.09 (s, 1H), 7.08 (s, 1H), 7.02 (d, J=7.5 Hz, 1H), 6.78 (s, 1H), 4.61 (bs, 3H), 3.85 (s, 3H), 3.82 (s, 3H), 3.48 (s, 3H), 3.43 (s, 3H), 3.20-3.10 (m, 6H), 2.74 (t, J=7.3 Hz, 2H), 2.63 (t, J=7.3 Hz, 2H), 2.56 (t, J=7.3 Hz, 2H), 1.90-1.70 (m, 6H), 1.60-1.40 (m, 39H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4, 171.3, 171.2, 155.9, 154.9, 153.2, 152.4, 147.7, 145.4, 141.7, 140.3, 139.8, 139.0, 134.1, 133.2, 128.3, 124.0, 123.5, 123.3, 120.9, 118.2, 115.5, 115.0, 112.1, 108.8, 106.5, 106.1, 104.0, 79.0, 60.7, 56.2, 55.7, 55.6, 40.3, 33.8, 33.7, 29.7, 28.4, 26.2, 26.0, 24.7, 24.5, 24.4.

MS (ESI) m/z: 1191 (M+23)⁺.

Rf: 0.19 (hexane:AcOEt, 1:1).

General procedure C (starting from 192) to afford 193 as a white solid (1.45 g, 90%).

¹H NMR (300 MHz, CD₃OD) δ 9.08 (d, J=8.5 Hz, 1H), 7.44 (d, J=1.7 Hz, 1H), 7.39 (d, J=8.0 Hz, 1H), 7.26 (dd, J=8.1, 1.7 Hz, 1H), 7.20-7.10 (m, 3H), 6.85 (s, 1H), 3.86 (s, 3H), 3.84 (s, 6H), 3.50 (s, 3H), 3.44 (s, 3H), 3.05-2.90 (m, 6H), 2.83 (t, J=7.3 Hz, 2H), 2.71 (t, J=7.3 Hz, 2H), 2.61 (t, J=7.3 Hz, 2H), 1.90-1.50 (m, 18H).

MS (ESI) m/z: 869 (M+1)⁺.

General procedure G (starting from 7-Hydroxy-8-bromo-isoquinoline) and chromatography on silica gel (CH₂Cl₂:EtOAc, 10:1) to afford 194 as a pale yellow solid (9 mg, 2%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=6.7 Hz, 1H), 7.63 (d, J=6.7 Hz, 1H), 7.18-7.05 (m, 5H), 6.96 (s, 1H), 6.62 (s, 1H), 4.72-4.55 (m, 2H), 3.84 (s, 3H), 3.44 (s, 3H), 1.5-1.40 (m, 12H).

Rf: 0.51 (CH₂Cl₂:EtOAc, 10:1).

General procedure C (starting from 38) to afford 195 as a pale yellow solid (654 mg, 83%).

¹H NMR (300 MHz, CD₃OD) δ 9.20-9.15 (m, 1H), 7.67 (s, 1H), 7.65-7.55 (m, 2H), 7.50-7.20 (m, 4H), 6.91 (d, J=8.4 Hz, 1H), 4.40-4.25 (m, 3H), 3.91 (d, J=3.8 Hz, 3H), 3.49 (s, 6H), 2.70-2.40 (m, 3H), 1.40-1.20 (m, 18H).

MS (ESI) m/z: 797 (M+1)⁺.

General procedure A (starting from 186) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 40:1) to afford 196 as a brown solid (25 mg, 62%).

¹H NMR (300 MHz, CDCl₃) δ 7.17 (s, 2H), 7.00 (s, 1H), 6.86 (s, 1H), 6.76 (s, 1H), 5.83 (bs, 1H), 5.78 (bs, 1H), 4.70 (t, J=6.4 Hz, 2H), 4.01 (s, 3H), 3.99 (s, 3H), 3.07 (t, J=6.7 Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 156.7, 147.9, 147.6, 146.6, 145.9, 141.7, 132.7, 132.6, 126.9, 119.4, 115.5, 114.7, 110.1, 108.3, 104.8, 104.2, 95.6, 56.6, 56.4, 42.7, 28.4.

MS (ESI) m/z: 380 (M+1)⁺.

Rf: 0.22 (CH₂Cl₂:MeOH, 40:1).

General procedure E (starting from 186 and 16 h of reaction time) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH from 50:1 to 10:1) to afford 197 as a beige solid (52 mg, 66%).

¹H NMR (300 MHz, CD₃OD) δ 8.77 (d, J=7.7 Hz, 1H), 7.28 (d, J=7.3 Hz, 1H), 7.20 (s, 1H), 6.81 (s, 1H), 6.63 (s, 1H), 4.75-4.55 (m, 2H), 3.96 (s, 3H), 3.70 (s, 3H), 3.49 (s, 3H), 1.50-1.40 (m, 6H), 1.35-1.25 (m, 6H).

¹³C NMR (75 MHz, CD₃OD+CDCl₃) δ 155.3, 154.0, 151.7, 148.3, 147.0, 146.4, 146.2, 143.7, 142.9, 133.1, 129.3, 129.2, 129.0, 126.8, 122.9, 121.8, 120.4, 119.3, 116.3, 114.0, 111.4, 108.3, 107.1, 104.9, 104.6, 103.1, 100.0, 99.7, 97.9, 76.4, 71.5, 60.5, 55.4, 22.4, 21.5, 21.3.

MS (ESI) m/z: 740 (M+23)⁺, 718 (M+1)⁺.

Rf: 0.14 (CH₂Cl₂:MeOH, 10:1).

General procedure A (starting from 197) and chromatography on silica gel Merck Si60 (230-400 mesh) (CH₂Cl₂:MeOH, 5:1) to afford 198 as a brown solid (15 mg, 42%).

¹H NMR (300 MHz, CD₃OD) δ 8.81 (d, J=7.5 Hz, 1H), 7.31 (d, J=7.5 Hz, 1H), 6.98 (s, 1H), 6.72 (s, 1H), 6.70 (s, 1H), 3.88 (s, 3H), 3.69 (s, 3H), 3.60 (s, 3H).

MS (ESI) m/z: 634 (M+1)⁺.

Rf: 0.22 (CH₂Cl₂:MeOH, 5:1).

General procedure C (starting from 41) to afford 199 as a pale yellow solid (537 mg, 80%).

¹H NMR (300 MHz, CD₃OD) δ 9.18 (d, J=7.5 Hz, 1H), 7.67 (s, 1H), 7.60-7.50 (m, 2H), 7.45-7.35 (m, 1H), 7.35-7.25 (m, 3H), 6.91 (d, J=8.0 Hz, 1H), 4.60-4.40 (m, 3H), 3.90 (d, J=2.5 Hz, 3H), 3.49 (s, 3H), 3.48 (s, 3H), 1.85-1.60 (m, 9H).

MS (ESI) m/z: 713 (M+1)⁺.

A suspension of 2 (100 mg, 0.18 mmol), Cs₂CO₃ (246 mg, 0.75 mmol) in anhydrous DMF (2 mL) was stirred at 23° C. under Argon atmosphere for 10 minutes, then 3-(BOC-amino)propyl bromide (180 mg, 0.75 mmol) was added and the mixture was heated at 50° C. overnight. The resulting solution was cooled to 23° C., quenched with H₂O, diluted with EtOAc (50 mL) and washed with H₂O (2×20 mL).

The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and the solvent removed under vaccum. The residue was purified by chromatography on silica gel (CH₂Cl₂:MeOH, 30:1) to afford 200 as a white solid (180 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.7 Hz, 1H), 7.36 (d, J=7.7 Hz, 1H), 7.20-7.10 (m, 3H), 6.97 (s, 1H), 6.93 (s, 1H), 6.70 (s, 1H), 5.50-5.40 (m, 2H), 4.97 (bs, 1H), 4.18 (t, J=6.4 Hz, 4H), 4.12 (t, J=5.8 Hz, 2H), 3.91 (s, 3H), 3.87 (s, 3H), 3.47 (s, 3H), 3.44 (s, 3H), 3.44-3.10 (m, 6H), 2.15-2.00 (m, 6H), 1.48 (s, 9H), 1.45 (s, 9H), 1.44 (s, 9H).

MS (ESI) m/z: 1023 (M+1)⁺.

Rf: 0.15 (hexane:AcOEt, 1:1).

Gerenal procedure C (starting from 200) to afford 201 as a white solid (110 mg, 85%).

¹H NMR (300 MHz, CD₃OD) δ 9.18 (d, J=7.5 Hz, 1H), 7.47 (d, J=7.5 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.28 (d, J=1.8 Hz, 1H), 7.23 (dd, J=8.0, 1.8 Hz, 1H), 7.10 (s, 1H), 7.07 (s, 1H), 6.80 (s, 1H), 4.28 (t, J=5.7 Hz, 4H), 4.21 (t, J=5.5 Hz, 2H), 3.91 (s, 3H), 3.88 (s, 6H), 3.47 (s, 3H), 3.46 (s, 3H), 3.30-2.25 (m, 4H), 3.19 (t, J=7.0 Hz, 2H), 2.30-2.15 (m, 6H).

MS (ESI) m/z: 701 (M+1)⁺.

Gerenal procedure C (starting from 203) to afford 202 as a pink solid (80 mg, 80%).

¹H NMR (300 MHz, CD₃OD) δ 9.02 (d, J=7.3 Hz, 1H), 7.40-7.30 (m, 3H), 7.30-7.15 (m, 3H), 6.96 (s, 1H), 6.77 (s, 1H), 4.27 (t, J=5.7 Hz, 4H), 4.35-4-15 (m, 2H), 3.90 (s, 3H), 3.47 (s, 3H), 3.46 (s, 3H), 3.40-3.20 (m, 6H), 2.20-2.10 (m, 6H), 1.58 (s, 9H), 1.48 (s, 9H), 1.44 (s, 9H).

MS (ESI) m/z: 671 (M+1)⁺.

A suspension of 3 (100 mg, 0.20 mmol), Cs₂CO₃ (293 mg, 0.90 mmol) in anhydrous DMF (2 mL) was stirred at 23° C. under argon atmosphere for 30 minutes, then 3-(BOC-amino)propyl bromide (214 mg, 0.90 mmol) was added and the mixture was heated at 40° C. for 4 hours. The resulting solution was cooled to 23° C., quenched with H₂O, diluted with EtOAc (50 mL) and washed with H₂O (2×20 mL).

The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and the solvent removed under vaccum. The residue was purified by chromatography on silica gel (CH₂Cl₂:MeOH, 30:1) to give 203 as a white solid (144 mg, 74%).

¹H NMR (300 MHz, CD₃OD) δ 9.24 (d, J=7.3 Hz, 1H), 7.25-7.10 (m, 4H), 7.08 (s, 1H), 7.04 (d, J=7.3 Hz, 1H), 6.93 (s, 1H), 6.72 (s, 1H), 5.50-5.40 (m, 3H), 4.30-4.10 (m, 6H), 3.87 (s, 3H), 3.47 (s, 3H), 3.46 (s, 3H), 3.30-2.10 (m, 6H), 2.30-2.15 (m, 6H).

MS (ESI) m/z: 971 (M+1)⁺.

Rf: 0.73 (CH₂Cl₂:MeOH, 30:1).

Gerenal procedure C (starting from 113) to afford 204 as a pale yellow solid (781 mg, 81%).

¹H NMR (300 MHz, CD₃OD) δ 9.17 (d, J=7.3 Hz, 1H), 7.60-7.25 (m, 21H), 7.17 (d, J=3.1 Hz, 1H), 6.89 (d, J=3.1 Hz, 1H), 4.80-4.60 (m, 3H), 3.94 (s, 3H), 3.60-3.40 (m, 12H).

MS (ESI) m/z: 941 (M+1)⁺.

General procedure D (starting from 3 and Boc-L-Leu-OH) and chromatography on silica gel (hexane:EtOAc, 3:2) to afford 205 as a yellow oil (100 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 9.24 (d, J=7.5 Hz, 1H), 7.45 (s, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.25-7.15 (m, 4H), 7.06 (d, J=7.5 Hz, 1H), 6.79 (d, J=7.1 Hz, 1H), 5.10-4.90 (m, 3H), 4.10-3.90 (m, 3H), 3.82 (s, 3H), 3.44 (s, 6H), 2.90-2.70 (m, 6H), 2.00-1.90 (m, 3H), 1.45 (s, 27H), 1.10-0.90 (m, 18H).

MS (ESI) m/z: 1161 (M+23)⁺.

Rf: 0.17 (hexane:EtOAc, 2:1).

General procedure C (starting from 205) to afford 206 as a white solid (66 mg, 85%).

¹H NMR (300 MHz, CD₃OD) δ 9.23 (d, J=7.7 Hz, 1H), 7.64 (s, 1H), 7.55-7.45 (m, 2H), 7.40-7.30 (m, 4H), 6.91 (s, 1H), 3.87 (s, 3H), 3.70-3.50 (m, 3H), 3.46 (s, 6H), 3.20-2.90 (m, 6H), 2.20-2.05 (m, 3H), 1.20-1.05 (m, 18H).

MS (ESI) m/z: 839 (M+1)⁺.

General procedure C (starting from 120) to afford 207 as a white solid (225 mg, 80%).

¹H NMR (300 MHz, CD₃OD) δ 9.09 (d, J=7.3 Hz, 1H), 7.60-7.30 (m, 18H), 7.20 (s, 1H), 7.13 (s, 1H), 7.12 (s, 1H), 6.87 (d, J=2.9 Hz, 1H), 4.76 (t, J=6.6 Hz, 2H), 4.62 (d, J=6.6 Hz, 1H), 4.00-3.85 (m, 6H), 3.70-3.35 (m, 12H).

MS (ESI) m/z: 971 (M+1)⁺.

General procedure D (starting from 3 and Boc-L-Ile-OH) and chromatography on silica gel (hexane:EtOAc, 2:1) to afford 208 as a yellow solid (537 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 9.26 (d, J=7.5 Hz, 1H), 7.42 (s, 1H), 7.35-7.15 (m, 5H), 7.09 (d, J=7.5 Hz, 1H), 6.79 (d, J=7.0 Hz, 1H), 5.10-5.05 (m, 3H), 4.60-4.55 (m, 3H), 3.79 (s, 3H), 3.43 (s, 6H), 2.20-2.05 (m, 3H), 1.70-1.60 (m, 3H), 1.49 (s, 9H), 1.47 (s, 9H), 1.45 (s, 9H), 1.40-1.20 (s, 6H), 1.15-0.90 (m, 18H).

MS (ESI) m/z: 1162 (M+23)⁺.

Rf: 0.45 (hexane:EtOAc, 2:1).

General procedure C (starting from 208) to afford 209 as a white solid (362 mg, 91%).

¹H NMR (300 MHz, CD₃OD) δ 9.24 (d, J=7.5 Hz, 1H), 7.67 (s, 1H), 7.60-7.50 (m, 2H), 7.45-7.30 (m, 4H), 6.92 (d, J=9.8 Hz, 1H), 4.40 (d, J=3.4 Hz, 1H), 4.37 (d, J=3.6 Hz, 1H), 4.33 (d, J=3.6 Hz, 1H), 3.88 (s, 3H), 3.49 (s, 3H), 3.48 (s, 6H), 2.30-2.10 (m, 3H), 1.90-1.70 (m, 3H), 1.60-1.40 (m, 3H), 1.30-1.00 (m, 18H).

MS (ESI) m/z: 839 (M+1)⁺.

General procedure D (starting from 3 and Alloc-Ala-OH) and chromatography on silica gel (CH₂Cl₂:MeOH, 80:1) to afford 210 as a white solid (29 mg, 74%).

¹H NMR (300 MHz, CDCl₃) δ 9.15-9.05 (m, 1H), 7.40-7.20 (m, 4H), 7.17 (d, J=6.5 Hz, 1H), 7.07 (s, 1H), 6.95-6.85 (m, 1H), 6.77 (d, J=5.8 Hz, 1H), 6.00-5.80 (m, 3H), 5.50-5.20 (m, 9H), 4.80-4.50 (m, 9H), 3.84 (d, J=2.9 Hz, 3H), 3.44 (s, 6H), 1.70-1.50 (m, 9H).

Rf: 0.14 (CH₂Cl₂:MeOH, 80:1).

To a suspension of 3 (20 mg, 0.04 mmol), Fmoc-Ala-OH (93 mg, 0.30 mmol) in CH₂Cl₂ anh. (2 mL) under Argon atmosphere at 0° C. was added HATU (114 mg, 0.30 mmol) and N-Methylmorpholine (0.053 mL, 0.48 mmol).

The mixture was stirred at 23° C. overnight. The resulting pale brown solution was diluted with CH₂Cl₂ (20 mL), washed with KHCO₃ 10% (20 mL), saturated aqueous solution of Na₂SO₄ (20 mL), and brine (20 mL).

The organic phase was dried over anhydrous Na₂SO₄ and the solvent removed under vaccum. The residue was purified by chromatography on silica gel (CH₂Cl₂:MeOH, 100:1) to give 211 as a white solid (32 mg, 84%).

¹H NMR (300 MHz, CDCl₃) δ 9.26 (d, J=7.7 Hz, 1H), 7.80-7.70 (m, 6H), 7.65-7.55 (m, 6H), 7.50-7.25 (m, 15H), 7.25-7.15 (m, 3H), 7.19 (d, J=6.9 Hz, 1H), 6.80-6.75 (m, 1H), 5.45-5.35 (m, 3H), 4.80-4.65 (m, 3H), 4.50-4.40 (m, 6H), 4.30-4.20 (m, 3H), 3.81 (s, 3H), 3.43 (s, 6H), 1.75-1.55 (m, 9H).

MS (ESI) m/z: 1401 (M+23)⁺.

Rf: 0.15 (CH₂Cl₂:MeOH, 100:1).

General procedure H (starting from 6,7-methylendioxi-3,4-dihidroisoquinoline) and chromatography on silica gel Merck-60 (230-400 mesh) (5:5:2 hexane-DCM-Et₂O) to provide 212 as a yellow solid (144 mg, 660%).

¹H NMR (300 MHz, CDCl₃) δ 7.10-6.95 (m, 3H), 6.90 (s, 1H), 6.74 (s, 1H), 6.62 (s, 1H), 6.58 (s, 1H), 5.89 (s, 2H), 4.80-4.50 (m, 4H), 3.82 (s, 3H), 3.41 (s, 3H), 3.08 (t, J=6.5 Hz, 2H), 1.50-1.25 (m, 12H).

MS (ESI) m/z: 588.2 (M+5)⁺.

Rf: 0.27 (hexane:EtOAc, 1:1).

General procedure E (starting from 212, reaction time 3 h) and chromatography on silica gel (hexane:EtOAc, 1:1) to give 213 (19 mg, 33%).

¹H NMR (300 MHz, CDCl₃) δ 9.23 (d, J=6.5 Hz, 1H), 7.20-6.90 (m, 7H), 6.63 (s, 1H), 6.00-5.95 (m, 2H), 4.80-4.50 (m, 2H), 3.83 (s, 3H), 3.43 (s, 3H), 1.50-1.20 (m, 12H).

MS (ESI) m/z: 582.2 (M+1)⁺.

Rf: 0.48 (hexane:EtOAc, 1:1).

General procedure M (starting from 4-dimethylaminophenyl boronic acid) and chromatography on silica gel (Hexane:EtOAc 3:1 to 2:1) to provide 214 (13 mg, 28%).

¹H NMR (300 MHz, CDCl₃) δ 7.35 (m, 2H), 6.89 (m, 3H), 6.76 (m, 3H), 4.78 (t, J=6.7 Hz, 2H), 4.54 (m, 2H), 3.44 (s, 3H), 3.33 (s, 3H), 3.08 (t, J=6.7 Hz, 2H), 2.98 (s, 6H), 1.37 (d, J=6.2 Hz, 6H), 1.36 (d, J=6.2 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.8, 150.5, 148.6, 147.2, 146.9, 146.6, 145.9, 136.2, 131.9, 128.5, 126.3, 123.0, 120.6, 115.5, 114.7, 113.7, 113.4, 110.8, 109.4, 105.2, 103.6, 71.5, 71.4, 55.6, 55.2, 42.4, 40.8, 29.3, 28.7, 22.1, 21.9.

MS (ESI) m/z: 583.5 (M+1)⁺.

Rf: 0.50 (hexane:EtOAc, 1:1).

General procedure H (starting from 6-isopropoxy-7-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (Hexane:CH₂Cl₂:Et₂O 5:5:2) to provide 215 as white solid (21 mg, 21%).

¹H NMR (300 MHz, CDCl₃) δ 7.73 (m, 4H), 7.44-7.35 (m, 6H), 7.20 (s, 1H), 6.94-6.80 (m, 3H), 6.75 (s, 1H), 6.71 (s, 1H), 6.67 (s, 1H), 5.23 (s, 2H), 4.84 (m, 1H), 4.68 (m, 1H), 4.56 (hp, J=6.0 Hz, 1H), 3.60 (s, 3H), 3.49 (s, 3H), 3.40 (s, 3H), 3.31 (s, 3H), 3.06 (m, 2H), 1.37 (d, J=6.0 Hz, 6H), 1.13 (s, 9H).

MS (ESI) m/z: 826.3 (M+1)⁺.

Rf: 0.40 (Hexane/CH₂Cl₂/Et₂O 5:5:2).

General procedure E (starting from 214, reaction time 6 h) and chromatography on silica gel (Hexane:EtOAc 1:1) to provide 216 (8 mg, 80%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.3 Hz, 1H), 7.39 (m, 2H), 7.30 (s, 1H), 7.08 (s, 1H), 7.00 (d, J=7.3 Hz, 1H), 6.96 (s, 1H), 6.84 (m, 3H), 4.69 (hp, J=6.2 Hz, 1H), 4.57 (hp, J=6.2 Hz, 1H), 3.48 (s, 3H), 3.47 (s, 3H), 2.92 (s, 3H), 1.43 (d, J=6.2 Hz, 6H), 1.40 (d, J=6.2 Hz, 6H).

MS (ESI) m/z: 867.4 (M+1)⁺.

Rf: 0.25 (Hexane:EtOAc 1:1).

General procedure M (starting from 3-nitrophenyl boronic acid) and chromatography on silica gel (Hexane:EtOAc 2:1) to provide 217 (33 mg, 67%) and LLSA-3,4-di(OiPr)-14(I) (10 mg, 20%).

¹H NMR (300 MHz, CDCl₃) δ 8.45 (s, 1H), 8.35 (d, J=8.1 Hz, 1H), 7.96 (d, J=7.7 Hz, 1H), 7.78 (dd, J=7.7, 8.1 Hz, 1H), 6.92 (s, 1H), 6.79 (s, 1H), 6.44 (s, 1H), 6.36 (s, 1H), 4.80 (dt, J=6.5, 6.3 Hz, 2H), 4.55 (m, 2H), 3.37 (s, 3H), 3.24 (s, 3H), 3.10 (t, J=6.5 Hz, 2H), 1.37 (d, J=6.1 Hz, 6H), 1.36 (d, J=6.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 148.9, 148.6, 148.0, 147.6, 146.7, 146.2, 138.2, 137.9, 136.1, 130.1, 127.8, 127.1, 126.4, 122.7, 119.3, 115.0, 114.3, 112.0, 109.7, 109.2, 104.6, 103.9, 71.6, 71.5, 55.6, 55.2, 42.5, 29.7, 22.0, 21.8.

MS (ESI) m/z: 585.4 (M+1)⁺.

Rf: 0.60 (Hexane:EtOAc 1:1).

General procedure M (starting from 3-thiopheneboronic acid) and chromatography on silica gel (Hexane:EtOAc 2:1) to provide 218 (18 mg, 39%).

¹H NMR (300 MHz, CDCl₃) δ 7.60 (dd, J=3.1, 5.0 Hz, 1H), 7.44 (dd, J=1.3, 3.1 Hz, 1H), 7.26 (dd, J=1.3, 5.0 Hz, 1H), 6.91 (s, 1H), 6.76 (s, 1H), 6.71 (s, 1H), 6.63 (s, 1H), 4.77 (m, 2H), 4.54 (m, 2H), 3.50 (s, 3H), 3.41 (s, 3H), 3.09 (t, J=6.6 Hz, 2H), 1.38 (d, J=6.0 Hz, 6H), 1.37 (d, J=6.0 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 148.7, 147.4, 147.0, 146.6, 145.9, 136.5, 135.5, 130.3, 128.8, 126.9, 126.3, 125.2, 120.0, 114.5, 113.9, 110.3, 108.9, 108.7, 104.4, 103.4, 71.4, 71.3, 55.4, 55.1, 42.4, 29.7, 28.6, 22.0, 21.8.

MS (ESI) m/z: 546.5 (M+1)⁺.

Rf: 0.65 (Hexane:EtOAc 1:1).

General procedure E (starting from 217, reaction time 5 h) and chromatography on silica gel (Hexane:EtOAc 2:1) to provide 219 (26 mg, quantitative).

¹H NMR (300 MHz, CDCl₃) δ 9.27 (d, J=7.4 Hz, 1H), 8.56 (m, 1H), 8.43 (m, 1H), 8.07 (d, J=7.7 Hz, 1H), 7.88 (dd, J=7.7, 8.0 Hz, 1H), 7.12 (s, 1H), 7.07 (d, J=7.4 Hz, 1H), 6.97 (s, 1H), 6.84 (s, 1H), 6.45 (s, 1H), 4.70 (hp, J=6.1 Hz, 1H), 4.57 (hp, J=6.1 Hz, 1H), 3.37 (s, 3H), 3.34 (s, 3H), 1.42 (d, J=6.1 Hz, 6H), 1.39 (d, J=6.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.3, 150.5, 149.0, 148.4, 148.9, 146.8, 146.7, 138.7, 138.5, 134.1, 130.3, 129.2, 127.0, 125.1, 123.2, 123.1, 118.3, 112.9, 110.8, 109.2, 108.5, 108.0, 105.1, 103.8, 71.6, 71.4, 55.5, 55.1, 29.7, 21.9, 21.8.

MS (ESI) m/z: 583.2 (M+1)⁺.

Rf: 0.60 (Hexane:EtOAc 1:1).

General procedure E (starting from 218, reaction time 5 h) and chromatography on silica gel (Hexane: EtOAc 2:1) to provide 220 (13 mg, 99%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.4 Hz, 1H), 7.70 (dd, J=3.0, 4.8 Hz, 1H), 7.56 (dd, J=1.3, 3.0 Hz, 1H), 7.34 (dd, J=1.3, 4.8 Hz, 1H), 7.16 (s, 1H), 7.09 (s, 1H), 7.02 (d, J=7.4 Hz, 1H), 6.96 (s, 1H), 6.71 (s, 1H), 4.70 (hp, J=6.2 Hz, 1H), 4.57 (d, J=6.2 Hz, 1H), 3.52 (s, 6H), 1.43 (d, J=6.2 Hz, 6H), 1.40 (d, J=6.2 Hz, 6H).

¹³C NMR (125 MHz, CDCl₃) δ 155.5, 150.2, 148.5, 147.9, 146.6, 135.8, 134.8, 130.8, 130.0, 127.2, 125.9, 124.7, 123.2, 118.9, 112.4, 110.3, 109.8, 108.1, 105.2, 104.9, 103.3, 71.4, 71.2, 55.4, 55.1, 29.7, 21.9, 21.8.

MS (ESI) m/z: 544.2 (M+1)⁺.

Rf: 0.65 (Hexane:EtOAc 1:1).

General procedure A (starting from 219) and chromatography on silica gel (CH₂Cl₂:MeOH 50:1) to provide 221 (14 mg, 88%).

¹H NMR (300 MHz, CDCl₃/CD₃OD) δ 9.14 (d, J=7.3 Hz, 1H), 8.52 (m, 1H), 8.46 (d, J=8.4 Hz, 1H), 8.08 (d, J=7.9 Hz, 1H), 7.92 (dd, J=7.9, 8.4 Hz, 1H), 7.14 (s, 1H), 7.06 (d, J=7.3 Hz, 1H), 6.90 (s, 1H), 6.82 (s, 1H), 6.42 (s, 1H), 3.37 (s, 3H), 3.36 (s, 3H).

MS (ESI) m/z: 499.4 (M+1)⁺.

Rf: 0.15 (CH₂Cl₂:MeOH 50:1).

A suspension of 3 (50 mg, 0.10 mmol) and Cs₂CO₃ (34 mg, 0.105 mmol) in anhydrous DMF (2 mL) under Argon atmosphere was heated at 40° C. for 30 minutes. Isopropylmagnesium bromide (0.014 mL, 0.15 mmol) was added dropwise via syringe to the reaction mixture. The resulting yellow suspension was stirred at 40° C. for 16 hours. The reaction mixture was cooled to 23° C. and evaporated in vacuo. The residue was disolved in CH₂Cl₂, filtered, and the solvent removed under vacuum.

The residue was purified by chromatography on silica gel (hexane:EtoAc 2:1 to 1:1) to afford 222 (30 mg, 51%).

¹H NMR (300 MHz, CDCl₃) δ 9.21 (d, J=7.3 Hz, 1H), 7.28-7.08 (m, 5H), 7.01 (d, J=7.3 Hz, 1H), 6.96 (s, 1H), 6.74 (s, 1H), 5.89 (s, 1H), 4.69 (hp, J=6.0 Hz, 1H), 4.57 (hp, J=6.2 Hz, 1H), 3.88 (s,3H), 3.46 (s, 3H), 3.45 (s, 3H), 1.43 (d, J=6.0 Hz, 6H), 1.40 (d, J=6.2 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 150.2, 148.5, 147.9, 147.3, 146.6, 145.7, 134.4, 129.5, 124.7, 123.1, 119.0, 115.2, 113.9, 112.3, 111.1, 110.5, 110.0, 107.8, 105.7, 105.6, 103.5, 71.4, 71.2, 56.2, 55.5, 55.2, 29.7, 21.9, 21.8.

MS (ESI) m/z: 584.2 (M+1)⁺.

Rf: 0.40 (Hexane/EtOAc 1:1).

General procedure A (starting from 220) and chromatography on silica gel (Hexane/EtOAc 1:1) to provide 223 (1.5 mg, 38%).

¹H NMR (300 MHz, CDCl₃/CD₃OD) δ 9.06 (d, J=7.4 Hz, 1H), 7.70 (dd, J=3.0, 4.8 Hz, 1H), 7.53 (dd, J=1.3, 3.0 Hz, 1H), 7.30 (dd, J=1.3, 4.8 Hz, 1H), 7.10 (s, 1H), 7.09 (s, 1H), 6.98 (d, J=7.4 Hz, 1H), 6.90 (s, 1H), 6.63 (s, 1H), 3.52 (s, 3H), 3.51 (s, 3H).

¹³C NMR (75 MHz, CDCl₃/CD₃OD) δ 156.4, 148.6, 148.1, 147.5, 147.0, 144.9, 136.1, 135.8, 130.9, 130.8, 127.6, 126.2, 125.7, 123.1, 118.7, 112.7, 111.4, 109.6, 107.9, 105.2, 105.0, 102.9, 102.8, 55.5, 55.2.

MS (ESI) m/z: 460.0 (M+1)⁺.

Rf: 0.20 (CH₂Cl₂:MeOH 50:1).

General procedure A (starting from 214) and chromatography on silica gel (CH₂Cl₂:MeOH 50:1 to 20:1) to provide 224 (11 mg, 50%).

¹H NMR (500 MHz, CDCl₃/CD₃OD) δ 7.43 (s, 1H), 7.31 (m, 2H), 6.91 (m, 2H), 6.81 (s, 1H), 6.72 (s, 1H), 6.70 (s, 1H), 4.67 (t, J=6.7 Hz, 2H), 3.43 (s, 3H), 3.33 (s, 3H), 3.02 (t, J=6.7 Hz, 2H), 2.96 (s, 6H).

¹³C NMR (125 MHz, CDCl₃/CD₃OD) δ 156.9, 151.1, 146.8, 146.6, 146.5, 146.4, 145.0, 137.5, 132.3, 129.7, 127.6, 123.6, 119.8, 115.4, 115.1, 114.0, 113.4, 110.5, 109.6, 105.3, 103.9, 55.7, 55.4, 42.8, 41.1, 28.7.

MS (ESI) m/z: 499.2 (M+1)⁺.

Rf: 0.15 (CH₂Cl₂:MeOH 40:1).

A suspension of 227 (63 mg, 0.080 mmol) and Cs₂CO₃ (29 mg, 0.088 mmol) in anhydrous DMF under Argon atmosphere at room temperature for 30 minutes. 4-methoxybenzyl chloride (0.088 mmol) was added dropwise via syringe to the reaction mixture. The resulting suspension was stirred at room temperature overnight. The progress of the reaction was followed by TLC (CH₂Cl₂/EtOAc 10:1). The reaction mixture was evaporated in vacuo. The residue was purified by chromatography on silica gel (CH₂Cl₂/EtOAc 10:1) to obtain 225 (9 mg, 12%).

¹H NMR (300 MHz, CDCl₃) δ 7.73 (m, 4H), 7.46-7.35 (m, 8H), 7.34 (s, 1H), 6.93-6.80 (m, 5H), 6.76 (s, 1H), 6.71 (s, 1H), 6.67 (s, 1H), 5.65 (s, 1H), 5.22 (s, 2H), 5.08 (s, 2H), 4.82 (m, 1H), 4.63 (m, 1H), 3.81 (s, 3H), 3.60 (s, 3H), 3.49 (s, 3H), 3.40 (s, 3H), 3.32 (s, 3H), 3.04 (m, 2H), 1.13 (s, 9H).

MS (ESI) m/z: 904.0 (M+1)⁺.

Rf: 0.65 (CH₂Cl₂/EtOAc 10:1).

A suspension of 227 (63 mg, 0.080 mmol) and Cs₂CO₃ (29 mg, 0.088 mmol) in anhydrous DMF under Argon atmosphere at room temperature for 30 minutes. 4-methoxybenzyl chloride (0.088 mmol) was added dropwise via syringe to the reaction mixture. The resulting suspension was stirred at room temperature overnight. The progress of the reaction was followed by TLC (CH₂Cl₂/EtOAc 10:1). The reaction mixture was evaporated in vacuo. The residue was purified by chromatography on silica gel (CH₂Cl₂/EtOAc 10:1) to obtain 226 (33 mg, 48%).

¹H NMR (300 MHz, CDCl₃) δ 7.40-7.33 (m, 8H), 7.25 (s, 1H), 7.21-7.00 (m, 3H), 6.92-6.88 (m, 4H), 6.77 (s, 1H), 6.69 (s, 1H), 6.64 (s, 1H), 5.22 (s, 2H), 5.19 (s, 2H), 5.07 (s, 2H), 4.82 (m, 1H), 4.64 (m, 1H), 3.85 (s, 3H), 3.82 (s, 3H), 3.81 (s, 3H), 3.49 (s, 3H), 3.36 (s, 3H), 3.27 (s, 3H), 3.05 (m, 2H).

MS (ESI) m/z: 786.0 (M+1)⁺.

Rf: 0.50 (CH₂Cl₂/EtOAc 10:1).

General procedure G (starting from 6-hydroxy-7-methoxy-3,4-dihydroisoquinoline and iodo-acetic acid 2-[4-(tert-butyl-diphenyl-silannyloxy)-3-methoxy-phenylethynyl]-4-methoxy-5-methoxymethoxy-phenyl ester) and chromatography on silica gel (Hexane:CH₂Cl₂:Et₂O 5:5:2) to provide 227 slightly impure (103 mg, 20%). To obtain a pure product, this compound was submitted to chromatography on LiChroprep® NH₂ (EtOAc) (24 mg, 5%).

¹H NMR (300 MHz, CDCl₃) δ 7.74 (m, 4H), 7.47-7.35 (m, 6H), 7.20 (s, 1H), 6.94-6.70 (m, 4H), 6.67 (m, 2H), 5.65 (s, 1H), 5.23 (s, 2H), 4.84 (m, 1H), 4.63 (m, 1H), 3.61 (s, 3H), 3.49 (s, 3H), 3.40 (s, 3H), 3.36 (s, 3H), 3.05 (m, 2H), 1.14 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 155.4, 151.1, 146.1, 145.9, 145.8, 145.7, 145.1, 144.9, 135.9, 135.1, 133.5, 133.4, 129.9, 128.3, 127.6, 127.4, 123.3, 120.3, 119.6, 115.1, 114.8, 114.1, 113.8, 111.9, 108.4, 105.3, 105.0, 95.5, 56.2, 55.7, 55.4, 55.3, 42.3, 29.6, 28.4, 26.7, 19.8. MS (APCI) m/z: 784.1 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂/MeOH 100:1).

A suspension of 227 (25 mg, 0.031 mmol), Boc-L-Ala-OH (12 mg, 0.063 mmol), EDC·HCl (12 mg, 0.063 mmol) and DMAP (0.8 mg, 0.0063 mmol) in CH₂Cl₂ anh. (2 mL) was stirred under argon atmosphere at room temperature for 2 h. The resulting solution was diluted with CH₂Cl₂ (20 mL), washed with water (20 mL) and saturated NaHCO₃ aqueous solution (20 mL).

The organic phase was dried over anhydrous sodium sulfate and the solvent removed under vaccum to give 228 as a white solid (30 mg, quant.).

¹H NMR (300 MHz, CDCl₃) δ 7.73 (m, 4H), 7.44-7.35 (m, 6H), 7.20 (s, 1H), 6.96-6.80 (m, 5H), 6.65 (s, 1H), 5.22 (s, 2H), 5.09 (m, 1H), 4.90 (m, 1H), 4.63 (m, 1H), 3.61 (s, 3H), 3.49 (s, 3H), 3.39 (s, 3H), 3.27 (s, 3H), 3.07 (m, 2H), 1.55 (d, J=7.2 Hz, 3H), 1.47 (s, 9H), 1.13 (s, 9H).

MS (ESI) m/z: 955.2 (M+1)⁺.

Rf: 0.65 (Hexane/EtOAc 1:1).

A suspension of 222 (25 mg, 0.043 mmol) and Cs₂CO₃ (21 mg, 0.064 mmol) in anhydrous DMF under Argon atmosphere was heated at 40° C. for 30 minutes. MeI (0.215 mmol) was added dropwise via syringe to the reaction mixture. The resulting yellow suspension was stirred at 40° C. for 3 hours. The progress of the reaction was followed by TLC (CH₂Cl₂/MeOH 8:0.2).

The reaction mixture was cooled to 23° C. and evaporated in vacuo. The residue was disolved in CH₂Cl₂, filtered, and the solvent removed under vacuum to afford 229 (22 mg, 85%).

¹H NMR (500 MHz, DMSO-d₆) δ 9.07 (d, J=7.4 Hz, 1H), 7.46 (s, 1H), 7.33 (d, J=7.4 Hz, 1H), 7.28 (d, J=8.2 Hz, 1H), 7.21 (d, J=2.5 Hz, 1H), 7.16 (dd, J=8.2, 2.5 Hz, 1H), 7.15 (s, 1H), 7.11 (s, 1H), 6.70 (s, 1H), 4.76 (hp, J=6.1 Hz, 1H), 4.69 (hp, J=6.1 Hz, 1H), 3.86 (s,3H), 3.75 (s, 3H), 3.34 (s, 3H), 3.32 (s, 3H), 1.31 (d, J=6.1 Hz, 3H), 1.30 (d, J=6.1 Hz, 3H), 1.27 (d, J=6.1 Hz, 3H), 1.26 (d, J=6.1 Hz, 3H).

¹³C NMR (125 MHz, CDCl₃) δ 155.6, 150.1, 149.8, 149.0, 148.5, 147.9, 146.6, 146.5, 134.4, 129.5, 128.3, 124.7, 124.1, 123.2, 118.9, 114.3, 112.3, 111.9, 110.9, 110.3, 109.9, 107.8, 105.6, 105.4, 103.4, 71.4, 71.1, 56.3, 56.1, 55.5, 55.2, 29.7, 21.9, 21.8.

MS (ESI) m/z: 598.4 (M+1)⁺.

Rf: 0.65 (CH₂Cl₂/MeOH 8:0.2).

This compound is a by-product of the synthesis of 222.

¹H NMR (300 MHz, CDCl₃) δ 9.26 (d, J=7.5 Hz, 1H), 7.28-7.11 (m, 4H), 7.10 (s, 1H), 7.07 (d, J=7.5 Hz, 1H), 6.97 (s, 1H), 6.74 (s, 1H), 3.99 (s, 6H), 3.92 (s, 3H), 3.88 (s, 3H), 3.47 (s, 3H), 3.46 (s, 3H).

MS (APCI) m/z: 842.2 (M+1)⁺.

Rf: 0.35 (CH₂Cl₂/MeOH 9:0.2).

A suspension of 222 (22 mg, 0.037 mmol) and AlCl₃ (12 mg, 0.092 mmol) in anhydrous CH₂Cl₂ (1 mL) was stirred at room temperature for 2′5 h under Argon atmosphere. CH₂Cl₂ and MeOH were added and then the solvent was evaporated under reduced pressure. The residue was purified by preparative TLC (CH₂Cl₂/MeOH 9:0.5) to provide 231 (3 mg, 15%).

¹H NMR (500 MHz, DMSO-d₆) δ 9.98 (s, 1H), 9.03 (d, J=7.4 Hz, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.25 (d, J=7.4 Hz, 1H), 7.21 (d, J=2.5 Hz, 1H), 7.20 (s, 1H), 7.16 (dd, J=8.2, 2.5 Hz, 1H), 7.14 (s, 1H), 7.10 (s, 1H), 6.69 (s, 1H), 4.69 (hp, J=6.1 Hz, 1H), 3.86 (s, 3H), 3.76 (s, 3H), 3.37 (s, 3H), 3.35 (s, 3H), 1.27 (d, J=6.1 Hz, 3H), 1.26 (d, J=6.1 Hz, 3H).

¹³C NMR (125 MHz, DMSO-d₆) δ 154.3, 149.9, 149.0, 148.6, 148.4, 147.5, 146.2, 146.1, 134.0, 128.6, 127.1, 124.7, 123.6, 122.0, 117.5, 114.6, 113.0, 112.6, 111.6, 110.7, 109.2, 106.7, 105.2, 103.2, 70.5, 56.0, 55.8, 54.8, 54.5, 29.0, 21.7, 21.6. MS (APCI) m/z: 556.1 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂/MeOH 9:0.2).

A suspension of 222 (22 mg, 0.037 mmol) and AlCl₃ (12 mg, 0.092 mmol) in anhydrous CH₂Cl₂ (1 mL) was stirred at room temperature for 2′5 h under Argon atmosphere. CH₂Cl₂ and MeOH were added and then the solvent was evaporated under reduced pressure. The residue was purified by preparative TLC (CH₂Cl₂/MeOH 9:0.5) to provide 232 (1 mg, 5%).

¹H NMR (500 MHz, DMSO-d₆) δ 9.88 (s, 1H), 9.06 (d, J=7.4 Hz, 1H), 7.46 (s, 1H), 7.30 (d, J=8.2 Hz, 1H), 7.29 (d, J=7.4 Hz, 1H), 7.21 (d, J=2.5 Hz, 1H), 7.16 (dd, J=8.2, 2.5 Hz, 1H), 7.10 (s, 1H), 6.88 (s, 1H), 6.68 (s, 1H), 4.76 (hp, J=6.1 Hz, 1H), 3.86 (s,3H), 3.76 (s, 3H), 3.36 (s, 3H), 3.32 (s, 3H), 1.31 (d, J=6.1 Hz, 3H), 1.30 (d, J=6.1 Hz, 3H).

MS (APCI) m/z: 556.1 (M+1)⁺.

Rf: 0.25 (CH₂Cl₂/MeOH 9:0.2).

A suspension of 222 (22 mg, 0.037 mmol) and AlCl₁₃ (12 mg, 0.092 mmol) in anhydrous CH₂Cl₂ (1 mL) was stirred at room temperature for 2′5 h under Argon atmosphere. CH₂Cl₂ and MeOH were added and then the solvent was evaporated under reduced pressure. The residue was purified by preparative TLC (CH₂Cl₂/MeOH 9:0.5) to provide 233 (5 mg, 26%).

¹H NMR (500 MHz, DMSO-d₆) δ 9.97 (s, 1H), 9.86 (s, 1H), 9.02 (d, J=7.4 Hz, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.23 (d, J=7.4 Hz, 1H), 7.21 (d, J=2.5 Hz, 1H), 7.20 (s, 1H), 7.16 (dd, J=8.2, 2.5 Hz, 1H), 7.09 (s, 1H), 6.87 (s, 1H), 6.67 (s, 1H), 3.86 (s, 3H), 3.76 (s, 3H), 3.36 (s, 3H), 3.35 (s, 3H).

¹³C NMR (125 MHz, DMSO-d₆) δ 154.3, 149.9, 149.0, 148.5, 148.3, 147.8, 146.3, 144.6, 134.0, 128.9, 127.3, 124.7, 123.6, 122.0, 117.4, 114.6, 113.1, 112.4, 111.5, 110.4, 108.2, 106.4, 105.6, 105.3, 103.7, 56.0, 55.8, 55.0, 54.5. MS (APCI) m/z: 514.1 (M+1)⁺.

Rf: 0.15 (CH₂Cl₂/MeOH 9:0.2).

General procedure E (starting from 228, reaction time 28 h) and chromatography on silica gel (hexane:EtOAc 2:1) to afford 234 as a white solid (24 mg, 81%).

¹H NMR (300 MHz, CDCl₃) δ 9.20 (d, J=7.3 Hz, 1H), 7.75 (d, J=6.7 Hz, 4H), 7.48-7.37 (m, 6H), 7.25 (s, 1H), 7.03 (s, 1H), 7.00 (d, J=7.3 Hz, 1H), 6.92 (s, 2H), 6.73 (s, 1H), 5.24 (s, 2H), 5.12 (m, 1H), 4.62 (m, 1H), 3.64 (s, 3H), 3.50 (s, 3H), 3.43 (s, 3H), 3.37 (s, 3H), 1.58 (d, J=7.0 Hz, 3H), 1.25 (s, 9H), 1.15 (s, 9H).

MS (APCI) m/z: 953.2 (M+1)⁺.

Rf: 0.25 (hexane/EtAcO, 2:1).

A suspension of 227 (63 mg, 0.080 mmol) and Cs₂CO₃ (29 mg, 0.088 mmol) in anhydrous DMF under Argon atmosphere at room temperature for 30 minutes. 4-methoxybenzyl chloride (0.088 mmol) was added dropwise via syringe to the reaction mixture. The resulting suspension was stirred at room temperature overnight. The progress of the reaction was followed by TLC (CH₂Cl₂/EtOAc 10:1).

The reaction mixture was evaporated in vacuo. The residue was purified by chromatography on silica gel (CH₂Cl₂/EtOAc 10:1) to obtain 235 (15 mg, 26%).

¹H NMR (300 MHz, CDCl₃) δ 7.35 (m, 2H), 7.22 (s, 1H), 7.14-7.06 (m, 2H), 6.98-6.88 (m, 3H), 6.78 (s, 1H), 6.72 (s, 1H), 6.69 (s, 1H), 5.76 (s, 1H), 5.22 (s, 2H), 5.09 (s, 2H), 4.82 (m, 1H), 4.64 (m, 1H), 3.86 (s, 3H), 3.80 (s, 3H), 3.49 (s, 3H), 3.47 (s, 3H), 3.37 (s, 3H), 3.05 (m, 2H).

MS (ESI) m/z: 664.4 (M+1)⁺.

Rf: 0.30 (CH₂Cl₂/EtOAc 10:1).

To a stirred solution of 189 (80 mg, 0.135 mmol), PdCl₂(PPh₃)₂ (5 mg, 0.006 mmol) and CuI (8 mg, 0.04 mmol) in 5:1 DMF-Et₃N (1.2 mL) under argon atmosphere at room temperature, trimethylsilylacetylene (0.04 mL, 0.27 mmol) was added via syringe. The reaction mixture was heated in a sealed tube at 90° C. for 5 hours, then the mixture was cooled to room temperature and quenched with water (10 mL). The mixture was diluted with ethyl acetate (20 mL) and washed with water (2×10 mL) and brine (2×10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure.

To a solution of the resulting oil in methanol/dichloromethane (3:2, 5 mL)), potassium carbonate (20 mg, 0.142 mmol) was added in portions at room temperature under argon atmosphere. After 2 hours, a saturated aqueous solution of NH₄Cl was added. The aqueous phase was extracted with CH₂Cl₂ (twice, 20 mL) and the organic phase was dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure and the residue was purified by chromatography on silica gel (Hexane/EtOAc 2:1) to obtain 236 (20 mg, 26%).

¹H NMR (300 MHz, CDCl₃) δ 8.24 (s, 1H), 8.16 (s, 1H), 6.91 (s, 1H), 6.80 (s, 1H), 4.71 (t, J=6.6 Hz, 2H), 4.60 (m, 2H), 3.92 (s, 6H), 3.64 (s, 1H), 3.06 (t, J=6.6 Hz, 2H), 1.41 (d, J=6.0 Hz, 12H).

¹³C NMR (75 MHz, CDCl₃) δ 155.3, 149.4, 148.6, 148.2, 147.2, 146.3, 141.7, 131.5, 126.7, 119.8, 114.8, 114.0, 110.4, 109.6, 105.3, 103.4, 82.9, 79.9, 71.7, 56.5, 56.3, 42.7, 28.5, 22.3, 22.1.

MS (ESI) m/z: 488.5 (M+1)⁺.

Rf: 0.45 (Hexane/EtOAc 2:1).

General Procedure H (starting from iodo-acetic acid 2-[4-(tert-butyl-diphenyl-silannyloxy)-3-methoxy-phenylethynyl]-4-methoxy-5-methoxymethoxy-phenyl ester and 6-Benzyloxy-7-methoxy-3,4-dihydroisoquinoline) and chromatography on silica gel (5:5:2 hexane-dichloromethane-ether) to afford 237 as a white solid (273 mg, 20%).

¹H NMR (300 MHz, CDCl₃) δ 7.74 (m, 4H), 7.45-7.30 (m, 14H), 7.20 (s, 1H), 6.94 (m, 1H), 6.84 (m, 1H), 6.74 (m, 1H), 6.68 (s, 1H), 5.23 (s, 2H), 5.16 (s, 2H), 4.82 (m, 1H), 4.64 (m, 1H), 3.61 (s, 3H), 3.50 (s, 3H), 3.41 (s, 3H), 3.35 (s, 3H), 3.03 (m, 2H), 1.14 (s, 9H).

¹³C NMR (75 MHz, CDCl₃) δ 155.5, 151.1, 148.1, 146.1, 145.9, 145.7, 145.0, 136.7, 135.7, 135.4, 135.1, 133.4, 129.9, 129.6, 128.6, 128.2, 128.0, 127.8, 127.7, 127.5, 127.2, 126.4, 123.3, 120.6, 120.3, 115.2, 115.1, 113.4, 112.0, 109.2, 105.4, 105.0, 95.6, 71.0, 56.2, 55.7, 55.4, 55.2, 42.4, 28.6, 26.7, 19.8.

MS (ESI) m/z: 875 (M+1)⁺.

Rf: 0.45 (hexane:dichloromethane:Et₂O, 5:5:2).

General Procedure C (starting from the corresponding protected lamellarin) to afford 238 as a yellow solid (5 mg, 15%).

¹H NMR (300 MHz, CD₃OD) δ 9.10 (d, J=7.3 Hz, 1H), 7.43 (d, J=7.3 Hz, 1H), 7.40 (d, J=1.9 Hz, 1H), 7.30 (dd, J=7.3, 1.9 Hz, 1H), 7.16 (s, 1H), 7.14-7.12 (m, 2H), 6.86 (s, 1H), 6.73 (d, J=7.3 Hz, 1H), 4.61 (br s, 1H), 3.84 (s, 3H), 3.49 (s, 6H), 1.63 (d, J=7.3 Hz, 1H).

MS (ESI) m/z: 572 (M+1)⁺.

To a solution of 234 (10 mg, 0.016 mmol) in anhydrous THF (1 mL) at −78° C., 0.03 mL of a 1M TBAF solution in THF and 0.7M acetic acid solution were added. The mixture was stirred for 15 min at −78° C. Sodium bicarbonate saturated solution was added (5 drops), the mixture was diluted with dichloromethane (2 mL), dried over sodium sulfate and concentrated to dryness. To the resulting residue, a cold 3.0M solution of HCl in ethyl acetate (1 mL) was added and the mixture was stirred at 0° C. for 1 hour. The reaction was concentrated and the residue was washed with hexane and dichloromethane to afford 239 as a white solid (4 mg, 63% yield).

¹H NMR (300 MHz, CD₃OD) δ 8.99 (d, J=7.8 Hz, 1H), 7.50 (d, J=2.0 Hz, 1H), 7.48 (d, J=8.8 Hz, 1H), 7.30 (dd, J=2.0, 7.8 Hz, 1H), 7.15-7.00 (m, 3H), 6.77 (s, 1H), 6.68 (d, J=7.3 Hz, 1H), 4.52 (q, J=7.3 Hz, 1H), 3.89 (s, 3H), 3.50 (s, 3H), 3.56 (s, 3H), 3.49 (s, 3H), 1.81 (d, J=7.3 Hz, 3H).

MS (ESI) m/z: 571 (M+1)⁺.

To a solution of the corresponding protected lamellarin (47 mg, 0.047 mmol) in anhydrous THF (5 mL) at −78° C., 0.14 mL of a 1M TBAF solution in THF and 0.7M acetic acid solution were added. The mixture was stired 15 min at −78° C. Sodium bicarbonate saturated solution was added (5 drops), the mixture was diluted with dichloromethane (2 mL), dried over sodium sulfate and concentrated to dryness. To the resulting residue, a cold 3.0M solution of HCl in ethyl acetate (1.3 mL) was added and the mixture was stirred at 0° C. for 1 hour. The reaction was concentrated and the residue was washed with hexane and dichloromethane to afford 240 as a white solid (4 mg, 14

¹H NMR (300 MHz, CD₃OD) δ 9.10 (d, J=7.5 Hz, 1H), 7.48 (m, 2H), 7.33 (m, 1H), 7.15 (m, 3H), 6.85 (s, 1H), 6.71 (d, J=7.5 Hz, 1H), 4.51 (m, 1H), 3.87 (s, 3H), 3.50 (s, 3H), 3.49 (s, 3H), 1.80 (d, J=7.3 Hz, 1H).

MS (ESI) m/z: 572 (M+1)⁺.

Example 2 Bioassays for Antitumoral Activity

The finality of these assays is to interrupt the growth of an “in vitro” tumor cell culture by means a continued exhibition of the cells to the sample to be testing.

Cell Lines NAME N° ATCC SPECIES TISSUE CHARACTERISTICS K-562 CCL-243 human leukemia erythroleukemia (pleural effusion) A-549 CCL-185 human lung lung carcinoma “NSCL” SK-MEL-28 HTB-72 human melanoma malignant melanoma HT-29 HTB-38 human colon colon adenocarcinoma LoVo CCL-229 human colon colon adenocarcinoma LoVo-Dox human colon colon adenocarcinoma (MDR) DU-145 HTB-81 human prostate prostate carcinoma, not androgen receptors LNCaP CRL- human prostate prostate 1740 adenocarcinoma, with androgen receptors SK-BR-3 HTB-30 human breast breast adenocarcinoma, Her2/neu+, (pleural effusion) IGROV human ovary ovary adenocarcinoma IGROV-ET human ovary ovary adenocarcinoma, characterized as ET-743 resistant cells HeLa CCL-2 human cervix cervix epitheloid carcinoma HeLa-APL CCL-3 human cervix cervix epitheloid carcinoma, character- ized as aplidine resistant cells PANC-1 CRL- human pancreas pancreatic epitheloid 1469 carcinoma Inhibition of Cell Growth by Calorimetric Assay.

A colorimetric type of assay, using sulforhodamine B (SRB) reaction has been adapted for a quantitative measurement of cell growth and viability (following the technique described by P. A. Skehan, et al., J. Natl. Cancer Inst. 1990, 82, 1107-1112).

This form of assay employs 96 well cell culture microplates of 9 mm diameter (T. Mosmann et al., J. of Immunological Methods 1983, 65, 55-63; G. T. Faircloth et al., J. of Tissue and Culture Methods 1988, 11, 201-205).

Most of the cell lines are obtained from American Type Culture Collection (ATCC) derived from different human cancer types.

The values for mean+/−SD of data from triplicate wells are calculated. Some parameters for cellular responses can be calculated: GI=growth inhibition, TGI=total growth inhibition (cytostatic effect) and LC=cell killing (cytotoxic effect).

Tables 1 illustrates data on the biological activity of the compounds of the present invention. TABLE 1 Activity data (Molar) DU-145 LN-caP SKOV-3 IGROV IGROV-ET SK-BR-3 MEL-28 H-MEC-1 1 GI50 8.24E−06 2.77E−07 1.37E−05 4.93E−07 4.44E−07 2.37E−06 2.24E−06 6.06E−07 TGI 1.18E−05 1.44E−06 1.88E−05 1.76E−06 1.20E−06 1.09E−05 5.04E−06 1.38E−06 LC50 1.70E−05 7.85E−06 1.88E−05 1.88E−05 1.88E−05 1.88E−05 1.13E−05 5.72E−06 2 GI50 6.40E−08 7.27E−08 1.40E−07 8.80E−07 1.98E−06 3.14E−07 3.02E−07 6.65E−07 TGI 1.40E−07 3.98E−07 5.10E−06 5.17E−06 8.12E−06 2.36E−06 3.66E−06 1.89E−05 LC50 1.89E−05 1.44E−06 1.89E−05 1.89E−05 1.89E−05 1.88E−05 1.89E−05 1.89E−05 3 GI50 1.90E−08 1.23E−07 5.37E−08 2.36E−07 2.14E−07 1.06E−07 8.81E−08 1.13E−07 TGI 7.97E−08 3.98E−07 2.40E−07 1.06E−06 1.16E−06 7.67E−07 1.77E−06 3.14E−06 LC50 2.00E−05 1.06E−06 1.58E−05 2.00E−05 2.00E−05 7.69E−06 1.13E−05 1.55E−05 4 GI50 2.18E−07 3.32E−07 4.02E−07 5.99E−07 3.38E−07 TGI 6.67E−07 7.85E−07 2.00E−06 1.64E−06 7.03E−07 LC50 2.14E−06 1.85E−06 1.13E−05 1.31E−05 1.47E−06 5 GI50 3.56E−08 2.23E−07 2.47E−07 2.12E−07 2.41E−07 6.29E−08 TGI 1.19E−07 1.05E−06 8.55E−07 1.06E−06 5.95E−07 2.49E−07 LC50 5.75E−07 3.62E−06 4.54E−06 6.09E−06 3.54E−06 1.12E−06 6 GI50 3.55E−07 5.78E−07 9.63E−07 1.39E−06 5.48E−07 3.86E−07 TGI 1.46E−06 7.96E−06 7.96E−06 7.96E−06 2.56E−06 1.74E−06 LC50 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7 GI50 9.93E−07 4.25E−07 1.23E−06 1.86E−06 1.14E−06 6.52E−07 TGI 2.31E−06 1.36E−06 2.67E−06 4.58E−06 2.61E−06 1.64E−06 LC50 5.41E−06 3.61E−06 5.79E−06 8.14E−06 5.98E−06 3.75E−06 8 GI50 7.03E−07 6.54E−07 4.45E−07 3.35E−06 9.15E−07 4.07E−07 TGI 2.60E−06 2.47E−06 2.17E−06 1.12E−05 3.04E−06 2.20E−06 LC50 9.43E−06 7.21E−06 1.12E−05 1.12E−05 9.19E−06 1.12E−05 9 GI50 5.43E−08 1.74E−07 1.82E−07 1.32E−07 1.74E−07 1.34E−07 TGI 1.69E−07 4.22E−07 9.09E−07 9.80E−07 3.74E−07 5.16E−07 LC50 6.15E−07 1.38E−06 3.00E−06 4.93E−07 8.06E−07 2.11E−06 11 GI50 3.53E−08 2.67E−07 5.53E−06 1.87E−07 2.00E−07 TGI 1.51E−07 1.14E−06 1.11E−05 1.20E−06 1.18E−06 LC50 2.78E−06 5.88E−06 1.11E−05 6.66E−06 6.18E−06 12 GI50 2.49E−07 1.42E−06 1.09E−05 6.98E−07 1.77E−06 TGI 1.43E−06 3.38E−06 1.09E−05 3.45E−06 5.98E−06 LC50 7.07E−06 8.01E−06 1.09E−05 1.09E−05 1.09E−05 13 GI50 8.32E−07 1.29E−06 1.96E−06 4.54E−08 3.11E−06 TGI 3.64E−06 4.79E−06 7.86E−06 1.13E−07 1.09E−05 LC50 1.24E−05 1.24E−05 1.24E−05 1.02E−05 1.24E−05 14 GI50 5.17E−07 4.90E−07 7.45E−07 4.24E−08 1.42E−06 TGI 2.19E−06 2.44E−06 4.97E−06 9.88E−08 1.40E−05 LC50 1.40E−05 1.40E−05 1.40E−05 1.40E−05 1.40E−05 A-549 K-562 PANC-1 HT-29 LOVO LOVO-DOX HELA HELA-APL 1 GI50 1.88E−06 8.15E−07 2.20E−06 1.88E−05 1.18E−05 8.01E−07 TGI 1.88E−06 2.11E−06 7.94E−06 1.88E−05 5.76E−05 7.13E−06 LC50 1.88E−06 2.97E−06 1.88E−05 1.88E−05 1.88E−05 1.88E−05 2 GI50 6.74E−07 1.17E−07 9.88E−07 8.31E−06 3.98E−07 6.04E−07 TGI 1.89E−05 2.25E−06 6.97E−06 1.89E−05 3.02E−06 3.83E−06 LC50 1.89E−05 1.89E−05 1.89E−05 1.89E−05 1.60E−05 1.89E−05 3 GI50 2.04E−08 8.69E−08 2.48E−07 5.33E−06 3.98E−07 4.20E−07 TGI 1.14E−06 7.91E−07 2.20E−06 1.90E−05 9.65E−07 1.67E−06 LC50 1.55E−05 8.15E−06 2.00E−05 2.00E−05 2.00E−06 2.00E−05 4 GI50 5.43E−07 3.96E−07 4.80E−07 2.50E−06 4.68E−07 2.00E−06 3.38E−07 2.38E−07 TGI 1.21E−06 9.37E−07 1.34E−06 6.03E−06 1.08E−06 5.95E−06 6.85E−07 6.81E−07 LC50 1.00E−05 4.44E−06 5.99E−06 1.46E−05 4.36E−06 1.78E−05 1.39E−06 1.95E−06 5 GI50 1.21E−07 7.70E−08 4.18E−07 1.70E−06 3.45E−07 4.09E−07 1.24E−07 3.10E−07 TGI 1.02E−06 1.10E−06 1.64E−06 3.24E−06 1.06E−06 1.10E−06 1.12E−06 1.71E−06 LC50 4.65E−06 7.40E−06 4.51E−06 6.16E−06 4.14E−06 3.40E−06 3.10E−06 5.55E−06 6 GI50 4.97E−07 4.82E−07 2.46E−06 7.96E−06 2.79E−06 3.84E−06 5.84E−07 3.41E−06 TGI 7.96E−06 5.38E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 LC50 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7.96E−06 7 GI50 9.69E−07 5.96E−07 1.36E−06 8.71E−07 2.00E−06 3.66E−06 8.30E−07 1.85E−06 TGI 2.96E−06 2.68E−06 3.31E−06 3.31E−06 5.05E−06 8.14E−06 1.91E−06 3.66E−06 LC50 8.14E−06 8.14E−06 8.04E−06 8.14E−06 8.14E−06 8.14E−06 4.42E−06 7.24E−06 8 GI50 7.44E−07 2.25E−07 2.31E−07 3.77E−07 1.48E−06 8.34E−06 2.76E−07 7.30E−06 TGI 6.34E−06 1.57E−06 8.65E−07 1.12E−05 4.68E−06 1.12E−05 1.41E−06 1.12E−05 LC50 1.12E−05 7.49E−06 1.12E−05 1.12E−05 1.12E−05 1.12E−05 7.92E−06 1.12E−05 9 GI50 3.23E−07 4.73E−08 2.00E−07 1.61E−06 2.43E−07 3.88E−07 1.07E−07 2.82E−07 TGI 1.15E−06 3.11E−07 1.56E−06 3.28E−06 5.84E−07 1.14E−06 4.59E−07 1.23E−06 LC50 6.76E−06 4.18E−06 4.91E−06 6.66E−06 2.30E−06 3.63E−06 4.06E−06 3.83E−06 11 GI50 1.28E−07 2.14E−07 3.84E−08 4.24E−08 1.11E−05 2.08E−07 5.23E−06 TGI 1.54E−06 1.76E−06 1.75E−07 1.16E−06 1.11E−05 1.35E−06 1.11E−05 LC50 1.11E−05 1.11E−05 8.23E−06 1.04E−05 1.11E−05 1.11E−05 1.11E−05 12 GI50 1.20E−06 1.62E−06 8.11E−08 1.83E−06 1.09E−05 1.66E−06 3.66E−06 TGI 6.64E−06 8.94E−06 8.41E−07 7.41E−06 1.09E−05 7.50E−06 1.09E−05 LG50 1.09E−05 1.09E−05 1.09E−05 1.09E−05 1.09E−05 1.09E−05 1.09E−05 13 GI50 9.06E−07 2.43E−06 3.22E−06 7.54E−07 6.75E−07 1.83E−06 2.60E−06 TGI 1.24E−05 1.24E−05 6.39E−06 2.35E−06 3.42E−06 9.63E−06 1.21E−05 LC50 1.24E−05 1.24E−05 1.24E−05 9.02E−06 1.24E−05 1.24E−05 1.24E−05 14 GI50 5.55E−07 1.18E−06 2.47E−06 5.97E−07 4.97E−07 1.83E−06 2.60E−06 TGI 1.40E−05 1.40E−05 7.26E−06 2.47E−06 1.45E−06 9.63E−06 1.21E−05 LC50 1.40E−05 1.40E−05 1.40E−05 1.40E−05 1.40E−05 1.24E−05 1.24E−05 DU-145 LN-caP SKOV-3 IGROV IGROV-ET SK-BR-3 MEL-28 H-MEC-1 15 GI50 6.85E−08 5.60E−08 8.62E−08 6.87E−06 5.61E−08 4.82E−08 TGI 3.08E−06 6.09E−07 2.03E−06 1.10E−05 6.35E−07 1.64E−06 LC50 1.10E−05 3.93E−06 9.76E−06 1.10E−05 5.84E−06 7.24E−06 16 GI50 5.27E−08 1.31E−07 2.26E−07 2.78E−07 2.07E−07 1.38E−07 TGI 2.98E−07 2.65E−07 4.78E−07 5.00E−07 4.11E−07 3.04E−07 LC50 2.80E−06 5.34E−07 1.59E−06 8.97E−07 8.16E−07 6.69E−07 17 GI50 4.87E−08 1.32E−07 1.05E−07 1.62E−07 8.03E−08 5.18E−08 TGI 2.28E−07 2.81E−07 4.05E−07 5.44E−07 2.86E−07 1.71E−07 LC50 2.30E−06 5.99E−07 4.93E−06 8.78E−06 1.22E−06 7.66E−07 18 GI50 5.05E−07 3.59E−07 1.22E−06 1.31E−06 2.00E−06 4.58E−07 5.24E−07 4.94E−07 TGI 1.41E−06 1.27E−06 2.82E−06 3.21E−06 5.14E−06 2.14E−06 1.44E−06 8.46E−07 LC50 7.04E−06 4.27E−06 6.51E−06 7.91E−06 1.23E−05 1.05E−05 4.43E−06 2.93E−06 19 GI50 5.32E−07 9.41E−08 7.58E−07 4.06E−07 7.61E−07 1.65E−07 2.73E−07 8.07E−07 TGI 2.16E−06 1.41E−06 2.31E−06 2.51E−06 2.54E−06 2.28E−06 1.53E−06 2.42E−06 LC50 6.85E−06 5.37E−06 6.57E−06 9.34E−06 1.13E−05 1.40E−05 4.99E−06 7.16E−06 20 GI50 5.00E−07 5.16E−07 5.93E−07 5.48E−08 2.24E−06 TGI 2.36E−06 7.46E−06 1.53E−05 2.38E−07 1.53E−05 LC50 1.53E−05 1.53E−05 1.53E−05 1.53E−05 1.53E−05 21 GI50 3.53E−07 1.90E−07 8.49E−07 3.15E−07 9.01E−07 3.42E−07 2.87E−07 2.81E−07 TGI 7.03E−07 3.83E−07 1.17E−05 6.64E−07 5.18E−06 5.99E−07 4.97E−07 5.09E−07 LC50 1.17E−05 7.75E−07 1.17E−05 9.02E−06 1.17E−05 1.17E−05 8.60E−07 9.22E−07 22 GI50 7.43E−07 5.64E−07 2.01E−07 7.87E−07 2.89E−06 2.57E−07 3.32E−07 8.56E−07 TGI 8.10E−06 1.82E−06 2.22E−06 8.54E−06 1.36E−05 1.81E−06 3.56E−06 2.91E−06 LC50 1.36E−05 1.36E−05 1.36E−05 1.36E−05 1.36E−05 7.16E−06 1.36E−05 1.04E−05 23 GI50 1.76E−07 1.68E−07 1.18E−08 3.30E−07 1.86E−06 3.91E−08 6.99E−08 2.19E−07 TGI 1.62E−06 5.34E−07 3.20E−07 1.92E−06 5.82E−06 3.03E−07 8.15E−07 6.87E−07 LC50 5.82E−06 2.34E−06 1.97E−06 5.82E−06 5.82E−06 2.85E−06 3.09E−06 2.87E−06 24 GI50 9.13E−07 9.34E−07 1.67E−06 1.23E−06 1.31E−06 6.53E−07 1.38E−06 8.06E−07 TGI 2.94E−06 1.91E−06 3.39E−06 4.82E−06 7.13E−06 1.93E−06 2.81E−06 2.20E−06 LC50 7.13E−06 3.89E−06 6.90E−06 7.13E−06 7.13E−06 5.58E−06 5.74E−06 5.97E−06 26 GI50 6.11E−07 7.11E−07 8.74E−07 5.04E−07 5.74E−07 9.66E−08 2.08E−06 7.98E−07 TGI 2.94E−06 4.30E−06 5.53E−06 2.30E−06 2.08E−06 1.09E−06 7.79E−06 1.95E−05 LC50 1.95E−05 1.47E−05 1.95E−05 1.30E−05 1.95E−05 1.36E−05 1.95E−05 1.95E−05 27 GI50 9.76E−07 1.15E−06 1.04E−05 8.81E−07 4.47E−06 TGI 3.95E−06 5.06E−06 1.52E−05 3.25E−06 9.35E−06 LC50 1.26E−05 1.52E−05 1.52E−05 1.41E−05 1.52E−05 28 GI50 3.15E−07 1.64E−06 1.14E−05 9.41E−07 2.24E−06 TGI 1.47E−06 3.97E−06 1.14E−05 4.73E−06 7.28E−06 LC50 7.32E−06 9.61E−06 1.14E−05 1.14E−05 1.14E−05 A-549 K-562 PANC-1 HT-29 LOVO LOVO-DOX HELA HELA-APL 15 GI50 5.41E−08 1.60E−06 1.54E−06 4.73E−08 1.68E−06 1.10E−05 5.79E−08 2.43E−06 TGI 1.46E−06 3.91E−06 5.26E−06 1.11E−07 4.96E−06 1.10E−05 1.41E−06 5.53E−06 LC50 1.10E−05 9.57E−06 1.10E−05 9.17E−06 1.10E−05 1.10E−05 7.81E−06 1.10E−05 16 GI50 1.78E−07 1.83E−07 2.41E−07 4.11E−07 1.96E−07 4.20E−07 1.93E−07 2.29E−07 TGI 5.02E−07 4.45E−07 7.56E−07 9.31E−07 3.47E−07 1.03E−06 3.68E−07 5.20E−07 LC50 3.05E−06 2.23E−06 3.34E−06 5.00E−06 6.12E−07 4.42E−06 7.03E−07 2.38E−06 17 GI50 2.07E−07 5.36E−08 2.06E−07 3.42E−07 2.16E−07 3.84E−07 1.09E−07 1.19E−07 TGI 6.63E−07 2.77E−07 1.52E−06 6.86E−07 4.20E−07 1.33E−06 3.59E−07 3.53E−07 LC50 8.48E−06 2.50E−06 8.53E−06 8.78E−06 8.16E−07 5.29E−06 8.78E−07 1.85E−06 18 GI50 1.61E−06 5.58E−07 9.92E−07 8.19E−07 6.64E−07 1.08E−06 TGI 4.00E−06 1.51E−06 3.70E−06 4.54E−06 2.37E−06 3.67E−06 LC50 9.99E−06 8.64E−06 1.23E−05 1.23E−05 1.23E−05 1.21E−05 19 GI50 6.05E−07 1.84E−06 2.45E−07 1.09E−07 7.22E−07 8.74E−07 TGI 2.13E−06 4.62E−06 2.42E−06 2.10E−06 2.33E−06 2.72E−06 LC50 6.32E−06 1.16E−05 9.50E−06 7.29E−06 6.60E−06 8.03E−06 20 GI50 5.89E−07 1.24E−06 3.58E−06 4.36E−07 5.25E−07 1.17E−06 2.09E−06 TGI 1.53E−05 1.53E−05 1.53E−05 8.83E−06 2.06E−06 1.53E−05 1.53E−05 LC50 1.53E−05 1.53E−05 1.53E−05 1.53E−05 1.53E−05 1.53E−05 1.53E−05 21 GI50 1.39E−06 2.86E−07 5.14E−07 5.12E−07 1.95E−06 4.97E−06 TGI 4.62E−06 5.82E−07 1.81E−06 1.17E−05 4.97E−06 1.17E−05 LC50 1.17E−05 2.39E−06 1.17E−05 1.17E−05 1.17E−05 1.17E−05 22 GI50 8.90E−07 6.70E−07 9.76E−07 8.79E−07 4.84E−07 6.59E−06 TGI 7.73E−06 4.77E−06 1.36E−05 1.36E−05 2.49E−06 1.36E−05 LC50 1.36E−05 1.36E−05 1.36E−05 1.36E−05 1.36E−05 1.36E−05 23 GI50 2.43E−07 2.48E−07 4.20E−07 2.90E−07 2.02E−07 1.61E−06 TGI 3.24E−06 1.01E−06 3.33E−06 5.82E−06 1.40E−06 4.70E−06 LC50 5.82E−06 3.36E−06 5.82E−06 5.82E−06 5.82E−06 5.82E−06 24 GI50 1.55E−06 6.80E−07 1.34E−06 7.13E−06 7.49E−07 8.91E−07 TGI 3.37E−06 1.91E−06 6.23E−06 7.13E−06 1.78E−06 3.66E−06 LC50 7.13E−06 5.30E−06 7.13E−06 7.13E−06 4.28E−06 7.13E−06 26 GI50 1.11E−06 7.85E−07 1.07E−06 9.17E−06 1.19E−06 8.67E−07 TGI 8.10E−06 3.41E−06 8.80E−06 1.95E−05 1.00E−05 7.63E−06 LC50 1.95E−05 1.61E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 27 GI50 5.49E−06 4.41E−06 1.44E−07 2.70E−07 1.52E−05 4.04E−06 1.30E−05 TGI 1.52E−05 1.30E−05 2.17E−06 2.17E−06 1.52E−05 9.47E−06 1.52E−05 LC50 1.52E−05 1.52E−05 1.52E−05 1.52E−05 1.52E−05 1.52E−05 1.52E−05 28 GI50 1.39E−06 1.91E−06 1.11E−07 6.34E−07 1.14E−05 2.76E−06 9.53E−06 TGI 1.14E−05 1.14E−05 1.20E−06 5.16E−06 1.14E−05 9.24E−06 1.14E−05 LC50 1.14E−05 1.14E−05 1.14E−05 1.14E−05 1.14E−05 1.14E−05 1.14E−05 DU-145 LN-caP SKOV-3 IGROV IGROV-ET SK-BR-3 MEL-28 H-MEC-1 29 GI50 1.98E−07 7.04E−08 1.79E−06 3.67E−07 5.65E−06 2.42E−07 1.55E−06 2.42E−08 TGI 4.40E−07 1.69E−07 3.90E−06 1.66E−06 7.35E−06 5.25E−07 3.17E−06 5.26E−08 LC50 3.90E−06 3.96E−07 7.35E−06 7.35E−06 7.35E−06 7.03E−06 6.45E−06 2.90E−07 30 GI50 5.38E−08 2.75E−07 3.48E−07 3.50E−07 4.50E−07 4.98E−07 2.51E−07 7.05E−08 TGI 2.65E−07 1.27E−06 1.24E−06 1.47E−06 1.83E−06 1.61E−06 1.08E−06 2.41E−07 LC50 3.84E−06 4.90E−06 5.36E−06 6.99E−06 6.63E−06 6.39E−06 4.42E−06 8.40E−07 31 GI50 3.01E−07 5.48E−07 5.24E−07 1.26E−06 1.39E−06 1.80E−06 4.82E−07 1.19E−07 TGI 8.61E−07 1.45E−06 4.22E−06 7.62E−06 4.00E−06 7.62E−06 7.62E−06 2.60E−07 LC50 7.62E−06 3.43E−06 7.62E−06 7.62E−06 7.62E−06 7.62E−06 7.62E−06 5.65E−07 32 GI50 1.11E−07 4.29E−07 1.86E−06 5.61E−07 6.66E−07 1.30E−06 1.35E−06 2.76E−07 TGI 3.09E−07 1.67E−06 3.67E−06 2.06E−06 3.01E−06 3.19E−06 2.87E−06 7.23E−07 LC50 8.57E−07 5.08E−06 7.27E−06 6.12E−06 8.76E−06 7.82E−06 6.13E−06 5.37E−06 33 GI50 1.86E−08 3.97E−08 3.91E−08 4.96E−08 7.81E−08 3.05E−08 4.11E−08 9.80E−09 TGI 4.60E−08 1.55E−07 2.28E−07 5.00E−07 5.89E−07 1.68E−07 1.98E−07 2.14E−08 LC50 3.67E−07 4.23E−07 5.03E−06 4.76E−06 8.83E−06 1.44E−06 2.48E−06 4.63E−08 34 GI50 1.96E−07 4.61E−07 1.27E−06 3.07E−07 2.07E−06 1.18E−06 1.36E−06 2.44E−07 TGI 5.88E−07 1.57E−06 2.91E−06 2.11E−06 4.63E−06 3.25E−06 2.76E−06 6.53E−07 LC50 6.50E−06 4.11E−06 6.65E−06 8.74E−06 9.10E−06 8.95E−06 5.58E−06 3.44E−06 35 GI50 3.21E−08 8.81E−08 1.13E−07 1.52E−07 1.92E−07 2.28E−07 5.46E−08 1.97E−08 TGI 8.81E−08 4.68E−07 9.32E−07 9.07E−07 8.55E−07 6.89E−07 9.15E−07 4.03E−08 LC50 5.76E−07 2.22E−06 3.83E−06 4.05E−06 5.45E−06 4.03E−06 2.77E−06 8.26E−08 36 GI50 1.08E−08 4.28E−08 3.92E−08 4.69E−08 7.65E−08 3.96E−08 3.19E−08 2.65E−09 TGI 3.83E−08 1.56E−07 2.97E−07 4.09E−07 5.40E−07 2.21E−07 1.81E−07 8.60E−09 LC50 6.68E−07 4.24E−07 5.89E−06 7.95E−06 8.78E−06 2.94E−06 2.70E−06 1.35E−06 37 GI50 6.77E−08 5.93E−07 1.62E−06 5.77E−07 3.59E−07 5.76E−08 6.23E−07 9.74E−08 TGI 1.55E−06 2.18E−06 3.58E−06 2.17E−06 2.11E−06 2.12E−06 2.68E−06 1.26E−06 LC50 5.62E−06 5.05E−06 7.87E−06 5.89E−06 8.04E−06 1.06E−05 7.95E−06 4.73E−06 38 GI50 5.12E−08 1.52E−07 6.84E−07 2.44E−07 1.29E−06 4.48E−07 1.65E−07 8.09E−08 TGI 9.11E−06 5.09E−07 7.42E−06 1.07E−06 4.51E−06 4.08E−06 5.20E−07 9.11E−07 LC50 9.11E−06 3.90E−06 9.11E−06 9.11E−06 9.11E−06 9.11E−06 9.11E−06 9.11E−06 39 GI50 1.80E−08 2.79E−08 6.34E−08 3.41E−08 5.50E−08 5.45E−08 3.39E−08 6.98E−09 TGI 1.42E−07 2.52E−07 8.47E−06 9.31E−07 3.69E−07 1.46E−06 2.95E−07 8.47E−06 LC50 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 40 GI50 1.62E−08 1.30E−07 2.84E−08 4.49E−08 8.52E−08 8.16E−07 4.12E−08 TGI 9.40E−08 4.47E−07 1.07E−07 3.91E−07 5.57E−07 3.25E−06 5.95E−07 LC50 9.48E−06 4.38E−06 9.48E−06 5.04E−06 9.48E−06 9.48E−06 9.48E−06 41 GI50 6.79E−07 2.10E−06 5.95E−06 9.17E−08 1.88E−07 1.62E−06 1.86E−06 TGI 9.87E−06 6.47E−06 9.87E−06 4.92E−07 8.52E−07 4.85E−06 4.19E−06 LC50 9.87E−06 9.87E−06 9.87E−06 4.70E−06 9.87E−06 9.87E−06 9.43E−06 42 GI50 3.65E−07 3.77E−07 1.24E−06 1.85E−07 2.38E−07 2.00E−06 1.23E−06 TGI 2.28E−06 1.41E−06 4.44E−06 5.60E−07 6.78E−07 4.01E−06 3.32E−06 LC50 9.46E−06 4.91E−06 9.46E−06 2.83E−06 6.62E−06 8.04E−06 8.94E−06 A-549 K-562 PANC-1 HT-29 LOVO LOVO-DOX 29 GI50 1.73E−06 4.81E−07 9.78E−07 1.84E−06 2.91E−06 7.35E−06 TGI 3.63E−06 3.46E−06 4.23E−06 5.08E−06 7.35E−06 7.35E−06 LC50 7.35E−06 7.35E−06 7.35E−06 7.35E−06 7.35E−06 7.35E−06 30 GI50 3.41E−07 2.04E−07 8.05E−07 2.81E−06 3.83E−07 6.83E−07 TGI 1.27E−06 1.10E−06 2.18E−06 5.06E−06 1.06E−06 1.93E−06 LC50 6.64E−06 8.60E−06 5.64E−06 8.60E−06 3.77E−06 5.22E−06 31 GI50 4.33E−07 4.10E−07 1.96E−06 7.62E−06 1.98E−06 1.49E−06 TGI 1.42E−06 2.47E−06 7.62E−06 7.62E−06 5.35E−06 7.62E−06 LC50 4.60E−06 7.62E−06 7.62E−06 7.62E−06 7.62E−06 7.62E−06 32 GI50 1.53E−06 6.73E−07 3.16E−07 2.66E−06 3.92E−07 6.71E−07 TGI 3.34E−06 2.43E−06 1.38E−06 5.30E−06 1.65E−06 2.42E−06 LC50 7.33E−06 8.31E−06 8.76E−06 8.76E−06 6.31E−06 8.25E−06 33 GI50 3.79E−08 5.96E−09 8.92E−08 1.34E−06 1.53E−07 1.98E−07 TGI 1.54E−07 9.18E−07 1.06E−06 3.42E−06 3.33E−07 5.58E−07 LC50 8.83E−07 4.03E−06 8.44E−06 8.74E−06 7.26E−07 4.33E−06 34 GI50 1.47E−06 1.04E−06 3.01E−07 1.16E−06 8.40E−07 9.10E−06 TGI 3.71E−06 2.61E−06 9.92E−07 5.39E−06 4.02E−06 9.10E−06 LC50 9.10E−06 6.62E−06 9.10E−06 9.10E−06 9.10E−06 9.10E−06 35 GI50 5.24E−08 8.08E−08 1.78E−07 1.18E−06 2.07E−07 2.17E−07 TGI 2.59E−07 9.75E−07 1.21E−06 2.44E−06 7.05E−07 7.37E−07 LC50 6.09E−06 4.82E−06 3.67E−06 5.06E−06 2.60E−06 2.80E−06 36 GI50 3.28E−08 9.48E−09 1.69E−07 1.51E−06 1.58E−07 1.45E−07 TGI 1.54E−07 1.88E−07 1.13E−06 4.08E−06 3.21E−07 5.25E−07 LC50 1.69E−06 5.70E−06 8.78E−06 8.78E−06 6.52E−07 8.78E−06 37 GI50 1.72E−06 1.24E−08 2.15E−06 9.12E−08 1.79E−06 2.10E−06 TGI 3.80E−06 1.97E−06 1.10E−05 2.00E−06 4.40E−06 4.60E−06 LC50 8.38E−06 7.53E−06 1.10E−05 1.09E−05 1.08E−05 1.01E−05 38 GI50 1.34E−06 4.13E−08 4.99E−07 3.24E−08 9.11E−06 9.11E−06 TGI 4.78E−06 1.15E−06 9.11E−06 9.11E−08 9.11E−06 9.11E−06 LC50 9.11E−06 9.11E−06 9.11E−06 9.11E−06 9.11E−06 9.11E−06 39 GI50 4.64E−08 3.59E−08 1.64E−07 3.04E−06 3.21E−07 3.23E−07 TGI 8.55E−07 1.08E−06 8.47E−06 8.47E−06 2.53E−06 1.81E−06 LC50 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 8.47E−06 40 GI50 3.16E−08 5.82E−08 2.54E−07 3.57E−06 2.23E−07 4.70E−07 TGI 2.70E−07 9.86E−07 3.27E−06 9.48E−06 8.84E−07 3.35E−06 LC50 9.48E−06 4.61E−06 9.48E−06 9.48E−06 6.41E−06 9.48E−06 41 GI50 2.42E−06 1.77E−06 2.48E−06 3.24E−06 3.31E−06 4.20E−06 TGI 8.01E−06 3.76E−06 9.87E−06 8.89E−06 9.87E−06 9.87E−06 LC50 9.87E−06 8.02E−06 9.87E−06 9.87E−06 9.87E−06 9.87E−06 42 GI50 7.35E−07 3.13E−07 7.32E−07 3.91E−06 4.63E−07 3.95E−07 TGI 2.99E−06 1.79E−06 4.06E−06 9.46E−06 3.63E−06 2.32E−06 LC50 9.46E−06 5.42E−06 9.46E−06 9.46E−06 9.46E−06 9.46E−06 DU-145 LN-caP SKOV-3 IGROV IGROV-ET SK-BR-3 MEL-28 H-MEC-1 43 GI50 5.74E−08 1.46E−07 4.18E−07 9.80E−08 2.12E−07 4.98E−07 TGI 3.01E−07 1.14E−06 2.14E−06 3.24E−06 1.10E−06 1.21E−05 LC50 1.21E−05 1.21E−05 8.94E−06 1.21E−05 7.90E−06 1.21E−05 44 GI50 4.85E−08 9.88E−08 1.06E−06 1.18E−06 1.22E−07 4.80E−07 TGI 2.73E−07 1.15E−06 2.32E−06 3.58E−06 8.66E−07 3.21E−06 LC50 7.60E−06 7.60E−06 5.06E−06 7.60E−06 7.60E−06 7.60E−06 45 GI50 1.05E−07 1.75E−07 1.57E−06 2.58E−06 4.14E−07 8.98E−07 TGI 5.49E−07 2.20E−06 4.12E−06 8.07E−06 1.58E−06 1.15E−05 LC50 1.15E−05 1.15E−05 1.09E−05 1.15E−05 1.15E−05 1.15E−05 46 GI50 3.80E−07 2.09E−06 3.16E−06 2.77E−07 4.99E−07 1.93E−06 1.22E−06 1.95E−07 TGI 2.19E−06 6.58E−06 9.57E−06 2.15E−06 3.03E−06 4.67E−06 9.57E−06 9.57E−06 LC50 8.54E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 46 GI50 2.76E−09 6.29E−10 3.20E−09 2.54E−09 2.87E−09 1.67E−09 2.21E−09 4.19E−10 TGI 5.71E−09 2.48E−09 9.57E−09 5.78E−09 6.57E−09 4.64E−09 4.17E−09 8.47E−10 LC50 9.57E−09 9.57E−09 9.57E−06 9.57E−07 9.57E−06 3.21E−08 7.87E−09 3.65E−09 47 GI50 1.08E−06 8.84E−08 7.64E−07 2.07E−06 2.29E−06 2.07E−06 TGI 4.61E−06 1.59E−06 1.03E−05 1.03E−05 6.01E−06 1.03E−05 LC50 1.03E−05 5.87E−06 1.03E−05 1.03E−05 1.03E−05 1.03E−05 53 GI50 4.53E−06 4.01E−06 1.38E−06 2.01E−05 1.99E−06 4.03E−06 TGI 1.23E−05 9.75E−06 2.01E−05 2.01E−05 9.49E−06 1.55E−05 LC50 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 54 GI50 2.88E−06 1.71E−06 8.84E−07 6.64E−06 1.05E−06 1.53E−06 TGI 7.16E−06 4.34E−06 4.02E−06 1.72E−05 4.31E−06 5.70E−06 LC50 1.72E−05 1.11E−05 1.33E−05 1.72E−05 1.72E−05 1.72E−05 57 GI50 3.89E−06 2.16E−06 8.47E−07 8.90E−07 3.17E−07 1.68E−06 TGI 1.22E−05 5.47E−06 5.88E−06 6.43E−06 5.24E−06 1.26E−05 LC50 1.95E−05 1.39E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 59 GI50 1.39E−06 1.37E−06 8.99E−07 6.32E−07 3.72E−07 1.62E−06 TGI 2.60E−06 2.63E−06 2.63E−06 2.04E−06 1.15E−06 3.04E−06 LC50 4.86E−06 5.06E−06 7.67E−06 5.98E−06 4.11E−06 5.67E−06 60 GI50 1.18E−06 5.50E−07 6.82E−07 2.56E−06 4.91E−07 1.17E−06 TGI 2.65E−06 1.79E−06 3.29E−06 6.16E−06 1.64E−06 4.14E−06 LC50 5.97E−06 5.25E−06 9.15E−06 9.15E−06 8.58E−06 9.15E−06 63 GI50 2.20E−06 1.13E−06 3.06E−06 9.75E−07 6.40E−07 1.57E−06 2.34E−06 2.72E−06 TGI 5.08E−06 3.47E−06 5.84E−06 3.86E−06 2.86E−06 4.80E−06 4.87E−06 5.25E−06 LC50 1.17E−05 9.61E−06 1.11E−05 1.46E−05 1.52E−05 1.47E−05 1.01E−05 1.01E−05 66 GI50 1.73E−06 8.90E−07 2.85E−06 1.35E−06 1.22E−06 4.34E−07 1.42E−06 1.52E−06 TGI 3.65E−06 1.89E−06 6.28E−06 3.02E−06 2.90E−06 1.82E−06 2.55E−06 2.73E−06 LC50 7.12E−06 4.03E−06 7.12E−06 6.75E−06 6.87E−06 7.12E−06 4.55E−06 4.93E−06 77 GI50 8.00E−07 7.51E−06 7.52E−06 5.92E−06 2.58E−07 3.53E−06 3.29E−06 2.71E−06 TGI 3.96E−06 1.17E−05 1.17E−05 8.84E−06 1.17E−06 1.17E−05 1.01E−05 1.17E−05 LC50 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 A-549 K-562 PANC-1 HT-29 LOVO LOVO-DOX HELA HELA-APL 43 GI50 1.90E−06 1.18E−07 1.49E−06 1.21E−05 6.35E−07 9.37E−07 TGI 1.21E−05 5.65E−07 6.57E−06 1.21E−05 3.72E−06 3.14E−06 LC50 1.21E−05 1.21E−05 1.21E−05 1.21E−05 1.21E−05 9.84E−06 44 GI50 5.44E−06 1.82E−07 1.17E−06 7.60E−06 3.74E−07 6.08E−07 TGI 7.60E−06 6.86E−07 4.35E−06 7.60E−06 3.18E−06 2.51E−06 LC50 7.60E−06 5.36E−06 7.60E−06 7.60E−06 7.60E−06 7.60E−06 45 GI50 4.35E−06 3.15E−07 2.03E−06 1.15E−05 7.71E−07 7.78E−07 TGI 1.15E−05 1.73E−06 9.75E−06 1.15E−05 6.71E−06 4.51E−06 LC50 1.15E−05 8.79E−06 1.15E−05 1.15E−05 1.15E−05 1.15E−05 46 GI50 1.31E−06 2.25E−06 1.34E−06 2.28E−06 4.23E−06 6.57E−06 TGI 9.57E−06 6.53E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 LC50 9.57E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 9.57E−06 46 GI50 2.33E−06 5.98E−10 2.94E−09 1.03E−08 3.24E−09 1.50E−08 TGI 9.57E−06 1.40E−09 1.15E−08 9.57E−06 9.32E−09 9.57E−08 LC50 9.57E−06 2.41E−09 9.57E−06 9.57E−06 9.57E−06 9.57E−06 47 GI50 3.26E−06 5.64E−07 3.86E−06 1.25E−06 3.97E−06 8.09E−06 5.37E−07 5.75E−06 TGI 1.03E−05 3.74E−06 1.03E−05 1.03E−05 1.03E−05 1.03E−05 4.79E−06 1.03E−05 LC50 1.03E−05 1.03E−05 1.03E−05 1.03E−05 1.03E−05 1.03E−05 1.03E−05 1.03E−05 53 GI50 5.66E−06 1.03E−05 1.03E−06 2.17E−06 1.11E−05 2.01E−05 5.90E−07 2.01E−05 TGI 2.01E−05 2.01E−05 9.67E−06 2.01E−05 2.01E−05 2.01E−05 1.23E−05 2.01E−05 LC50 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 54 GI50 2.85E−06 3.76E−06 7.11E−07 1.52E−06 3.40E−06 1.25E−05 6.16E−07 3.83E−06 TGI 1.51E−05 1.42E−05 2.59E−06 1.72E−05 8.94E−06 1.72E−05 2.71E−06 1.72E−05 LC50 1.72E−05 1.72E−05 1.72E−05 1.72E−05 1.72E−05 1.72E−05 1.72E−05 1.72E−05 57 GI50 1.29E−06 2.69E−06 6.11E−07 3.49E−06 7.17E−07 8.28E−07 6.85E−07 3.68E−06 TGI 1.95E−05 1.95E−05 9.70E−06 1.95E−05 5.30E−06 6.91E−06 5.67E−06 1.10E−05 LC50 1.95E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 1.95E−05 59 GI50 1.70E−06 1.64E−06 1.10E−06 1.97E−06 1.13E−06 5.28E−07 1.37E−06 1.88E−06 TGI 3.29E−06 3.45E−06 3.48E−06 3.70E−06 2.53E−06 1.60E−06 2.98E−06 3.90E−06 LC50 6.36E−06 7.27E−06 8.81E−06 6.98E−06 5.67E−06 4.56E−06 6.44E−06 8.12E−06 60 GI50 1.62E−06 6.97E−07 7.47E−07 2.42E−06 1.61E−06 2.82E−06 1.02E−06 3.49E−06 TGI 4.63E−06 2.19E−06 4.40E−06 5.88E−06 3.58E−06 9.15E−06 2.51E−06 9.15E−06 LC50 9.15E−06 6.87E−06 9.15E−06 9.15E−06 7.94E−06 9.15E−06 6.13E−06 9.15E−06 63 GI50 2.84E−06 2.90E−06 1.93E−06 2.92E−06 1.46E−06 9.09E−07 TGI 5.57E−06 6.75E−06 5.05E−06 5.70E−06 3.68E−06 2.95E−06 LC50 1.09E−05 1.52E−05 1.32E−05 1.11E−05 9.11E−06 9.85E−06 66 GI50 1.86E−06 9.82E−07 1.95E−06 2.77E−06 2.19E−06 1.91E−06 TGI 3.46E−06 2.80E−06 7.12E−06 6.77E−06 7.12E−06 5.56E−06 LC50 6.43E−06 7.12E−06 7.12E−06 7.12E−06 7.12E−06 7.12E−06 77 GI50 6.69E−06 2.13E−06 4.46E−06 1.17E−05 1.17E−05 1.02E−05 TGI 1.17E−05 6.15E−06 1.17E−05 1.17E−05 1.17E−05 1.17E−05 LC50 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 DU-145 LN-caP SKOV-3 IGROV IGROV-ET SK-BR-3 MEL-28 H-MEC-1 84 GI50 3.41E−06 7.10E−07 4.58E−06 2.41E−06 4.26E−06 4.60E−07 2.07E−06 6.82E−07 TGI 8.36E−06 2.01E−06 9.90E−06 6.28E−06 8.25E−06 9.50E−07 3.86E−06 1.86E−06 LC50 9.90E−06 5.21E−06 9.90E−06 9.90E−06 9.90E−06 9.90E−06 7.21E−06 5.96E−06 92 GI50 1.49E−05 1.67E−07 7.61E−06 3.13E−06 6.85E−06 4.35E−06 4.75E−06 1.42E−05 TGI 1.49E−05 7.10E−07 1.49E−05 6.64E−06 1.49E−05 1.16E−05 1.01E−05 1.49E−05 LC50 1.49E−05 5.09E−06 1.49E−05 1.41E−05 1.49E−05 1.49E−05 1.49E−05 1.49E−05 93 GI50 1.28E−05 4.08E−07 6.26E−06 3.37E−06 1.17E−05 4.95E−06 4.57E−06 1.28E−05 TGI 1.28E−05 1.27E−06 1.28E−05 7.08E−06 1.28E−05 1.28E−05 1.28E−05 1.28E−05 LC50 1.28E−05 7.08E−06 1.28E−05 1.28E−05 1.28E−05 1.28E−05 1.28E−05 1.28E−05 94 GI50 1.31E−05 6.73E−07 8.48E−06 1.83E−06 1.59E−06 3.35E−06 2.45E−06 1.59E−05 TGI 1.59E−05 2.87E−06 1.59E−05 5.90E−06 1.59E−05 1.24E−05 7.31E−06 1.59E−05 LC50 1.59E−05 1.25E−05 1.59E−05 1.59E−05 1.59E−05 1.59E−05 1.59E−05 1.59E−05 96 GI50 4.26E−06 4.20E−06 1.20E−06 2.69E−06 4.17E−07 2.14E−06 3.81E−07 TGI 9.20E−06 9.20E−06 3.01E−06 5.60E−06 1.86E−06 4.43E−06 9.20E−06 LC50 9.20E−06 9.20E−06 7.54E−06 9.20E−06 5.47E−06 9.18E−06 9.20E−06 97 GI50 4.13E−06 6.01E−06 1.86E−07 6.78E−07 1.23E−06 2.62E−07 3.37E−08 TGI 9.59E−06 9.59E−06 1.94E−06 3.33E−06 3.58E−06 4.71E−06 9.59E−08 LC50 9.59E−06 9.59E−06 7.54E−06 9.59E−06 9.59E−06 9.59E−06 9.59E−06 99 GI50 9.44E−06 4.46E−06 9.44E−06 3.07E−06 9.44E−06 9.44E−06 9.44E−06 4.06E−06 TGI 9.44E−06 8.19E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 5.48E−06 LC50 9.44E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 7.42E−06 101 GI50 1.31E−05 8.25E−06 7.91E−07 1.19E−05 1.31E−05 1.31E−05 1.31E−05 4.66E−06 TGI 1.31E−05 1.31E−05 2.34E−06 1.31E−05 1.31E−05 1.31E−05 1.31E−05 6.99E−06 LC50 1.31E−05 1.31E−05 5.71E−06 1.31E−05 1.31E−05 1.31E−05 1.31E−05 1.05E−05 109 GI50 2.99E−06 1.16E−06 2.49E−06 1.80E−06 9.67E−06 3.45E−06 8.85E−07 9.17E−07 TGI 1.54E−05 1.99E−06 1.18E−05 6.08E−06 1.99E−05 8.10E−06 2.57E−06 1.99E−05 LC50 1.99E−05 1.99E−05 1.99E−05 1.99E−05 1.99E−05 1.90E−05 1.08E−05 1.99E−05 115 GI50 1.63E−06 6.70E−07 1.10E−06 1.07E−06 1.70E−06 1.87E−06 1.45E−06 2.13E−07 TGI 4.29E−06 1.67E−06 2.38E−06 2.58E−06 5.38E−06 3.75E−06 2.65E−06 4.52E−07 LC50 7.78E−06 3.87E−06 5.13E−06 6.26E−06 7.78E−06 7.53E−06 4.83E−06 2.96E−06 118 GI50 1.60E−06 9.87E−07 2.02E−06 9.70E−07 8.76E−07 6.39E−07 1.65E−06 1.45E−06 TGI 3.00E−06 2.16E−06 4.08E−06 2.30E−06 2.33E−06 2.04E−06 3.00E−06 3.06E−06 LC50 5.64E−06 4.76E−06 8.22E−06 5.47E−06 6.17E−06 6.25E−06 5.49E−06 6.45E−06 126 GI50 2.53E−06 1.25E−06 3.48E−06 2.26E−06 3.88E−06 2.11E−06 2.93E−06 7.46E−07 TGI 6.49E−06 2.76E−06 6.98E−06 5.32E−06 7.87E−06 5.36E−06 5.27E−06 2.25E−06 LC50 1.29E−05 6.04E−06 1.29E−05 1.26E−05 1.29E−05 1.29E−06 9.48E−06 6.46E−06 128 GI50 1.33E−06 1.26E−06 6.39E−07 6.89E−07 1.57E−06 1.53E−06 TGI 2.48E−06 2.69E−06 1.98E−06 2.26E−06 3.03E−06 3.16E−06 LC50 4.62E−06 5.71E−06 5.70E−06 7.59E−06 5.86E−06 6.53E−06 134 GI50 2.12E−06 4.58E−07 7.39E−07 9.43E−07 4.82E−07 2.10E−06 1.73E−06 2.13E−06 TGI 4.47E−06 1.34E−06 1.85E−06 2.48E−06 2.14E−06 4.34E−06 3.23E−06 6.51E−06 LC50 8.81E−06 4.16E−06 4.26E−06 6.50E−06 7.58E−06 8.81E−06 6.04E−06 8.81E−06 A-549 K-562 PANC-1 HT-29 LOVO LOVO-DOX 84 GI50 2.66E−06 7.31E−07 5.01E−07 2.34E−06 4.48E−06 9.90E−06 TGI 4.91E−06 3.30E−06 5.47E−06 9.90E−06 9.90E−06 9.90E−06 LC50 9.03E−06 9.90E−06 9.90E−06 9.90E−06 9.90E−06 9.90E−06 92 GI50 1.49E−05 2.23E−06 5.42E−06 1.49E−05 7.52E−06 2.14E−06 TGI 1.49E−05 1.38E−05 1.49E−05 1.49E−05 1.49E−05 5.96E−06 LC50 1.49E−05 1.49E−05 1.49E−05 1.49E−05 1.49E−05 1.49E−05 93 GI50 8.64E−06 1.62E−06 1.28E−05 1.28E−05 1.28E−05 3.39E−06 TGI 1.28E−05 2.65E−06 1.28E−05 1.28E−05 1.28E−05 1.17E−05 LC50 1.28E−05 4.36E−06 1.28E−05 1.28E−05 1.28E−05 1.28E−05 94 GI50 7.19E−06 5.26E−07 2.12E−06 1.55E−05 5.70E−06 2.84E−06 TGI 1.59E−05 1.82E−06 1.59E−05 1.59E−05 1.59E−05 8.25E−06 LC50 1.59E−05 4.32E−06 1.59E−05 1.59E−05 1.59E−05 1.59E−05 96 GI50 4.97E−06 1.63E−06 1.68E−06 9.20E−06 2.28E−06 5.63E−07 TGI 9.20E−06 4.43E−06 5.65E−06 9.20E−06 9.20E−06 2.07E−06 LC50 9.20E−06 9.20E−06 9.20E−06 9.20E−06 9.20E−06 9.20E−06 97 GI50 9.59E−06 3.20E−06 4.47E−06 9.59E−06 5.28E−06 5.29E−06 TGI 9.59E−06 6.86E−06 9.59E−06 9.59E−06 9.59E−06 9.59E−06 LC50 9.59E−06 9.59E−06 9.59E−06 9.59E−06 9.59E−06 9.59E−06 99 GI50 9.44E−06 9.72E−07 9.44E−06 9.44E−06 9.44E−06 9.44E−06 TGI 9.44E−06 1.48E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 LC50 9.44E−06 2.26E−06 9.44E−06 9.44E−06 9.44E−06 9.44E−06 101 GI50 1.31E−05 2.19E−06 1.31E−05 1.31E−05 1.31E−05 1.31E−05 TGI 1.31E−05 3.53E−06 1.31E−05 1.31E−05 1.31E−05 1.31E−05 LC50 1.31E−05 5.73E−06 1.31E−05 1.31E−05 1.31E−05 1.31E−05 109 GI50 5.94E−06 4.25E−07 5.70E−06 4.81E−06 9.17E−07 5.64E−06 TGI 1.41E−05 1.99E−05 1.99E−05 1.99E−05 4.07E−06 1.35E−05 LG50 1.99E−05 1.99E−05 1.99E−05 1.99E−05 1.99E−05 1.99E−05 115 GI50 1.65E−06 5.36E−07 2.47E−06 2.92E−06 7.03E−07 1.76E−06 TGI 3.11E−06 2.47E−06 7.78E−06 6.46E−06 1.82E−06 4.78E−06 LC50 5.87E−06 7.78E−06 7.78E−06 7.78E−06 4.41E−06 7.78E−06 118 GI50 1.80E−06 1.17E−06 1.08E−06 1.92E−06 1.35E−06 4.50E−07 TGI 3.55E−06 2.77E−06 2.40E−06 3.56E−06 2.70E−06 1.30E−06 LC50 6.99E−06 6.57E−06 5.35E−06 6.59E−06 5.43E−06 3.86E−06 126 GI50 2.98E−06 2.15E−06 2.19E−06 3.00E−06 2.56E−06 1.29E−05 TGI 5.74E−06 4.32E−06 6.57E−06 9.22E−06 5.68E−06 1.29E−05 LC50 2.25E−06 8.69E−06 1.29E−05 1.29E−05 1.26E−05 1.29E−05 128 GI50 1.61E−06 1.11E−06 8.88E−07 1.95E−06 8.54E−07 4.03E−07 TGI 3.23E−06 2.98E−06 2.31E−06 3.54E−06 1.92E−06 1.49E−06 LC50 6.45E−06 8.01E−06 5.99E−06 6.44E−06 4.30E−06 4.66E−06 134 GI50 2.55E−06 5.71E−07 2.17E−06 2.85E−06 2.05E−06 5.19E−07 TGI 4.93E−06 4.93E−06 4.53E−06 8.81E−06 4.21E−06 1.99E−06 LC50 8.81E−06 8.81E−06 8.81E−06 8.81E−06 8.67E−06 7.79E−06 DU-145 LN-caP IGROV IGROV-ET SK-BR-3 MEL-28 A-549 K-562 135 GI50 1.60E−06 7.83E−07 2.10E−06 5.86E−07 5.79E−07 1.67E−06 3.34E−06 1.07E−06 TGI 3.78E−06 2.02E−06 4.68E−06 1.98E−06 2.61E−06 5.00E−06 8.76E−06 6.82E−06 LC50 8.76E−06 4.91E−06 8.76E−06 6.33E−06 8.76E−06 8.76E−06 8.76E−06 8.76E−06 169 GI50 1.51E−08 3.73E−08 4.71E−08 4.18E−08 3.87E−08 4.64E−08 6.46E−08 6.80E−08 TGI 9.01E−08 1.83E−07 1.92E−06 3.01E−07 1.32E−07 5.29E−07 1.14E−06 3.21E−06 LG50 1.05E−08 1.24E−06 1.05E−05 1.05E−05 1.05E−05 1.05E−05 9.74E−06 1.05E−05 173 GI50 4.01E−08 8.15E−08 1.53E−07 1.45E−07 1.52E−07 2.60E−07 1.11E−07 TGI 1.34E−07 2.68E−07 8.09E−07 4.28E−07 6.04E−06 6.04E−06 6.04E−06 LC50 7.49E−07 2.55E−06 6.04E−06 6.04E−06 6.04E−06 6.04E−06 6.04E−06 180 GI50 3.59E−08 5.30E−08 5.30E−08 6.91E−08 1.81E−08 1.34E−08 2.01E−08 4.00E−08 TGI 8.42E−08 2.27E−07 3.19E−07 5.31E−07 5.41E−08 1.26E−07 2.33E−07 1.53E−06 LC50 1.11E−05 6.15E−07 1.11E−05 1.11E−05 1.19E−06 1.11E−05 7.26E−06 1.11E−05 181 GI50 2.49E−08 5.93E−08 3.80E−08 5.00E−08 2.06E−08 1.68E−08 1.98E−08 3.28E−08 TGI 5.94E−08 2.37E−07 2.31E−07 3.41E−07 5.92E−08 9.56E−08 2.09E−07 4.44E−07 LC50 1.05E−05 6.78E−07 3.20E−06 1.05E−05 8.62E−07 2.14E−06 8.18E−06 7.14E−06 182 GI50 1.08E−06 1.75E−06 2.86E−06 3.16E−06 3.65E−07 1.08E−06 9.58E−06 3.16E−07 TGI 2.13E−05 2.13E−05 2.13E−05 2.13E−05 1.25E−06 2.13E−05 2.13E−05 4.85E−06 LC50 2.13E−05 2.13E−05 2.13E−05 2.13E−05 1.20E−05 2.13E−05 2.13E−05 2.13E−05 188 GI50 3.19E−06 3.30E−06 5.37E−06 2.51E−06 3.60E−06 1.14E−06 1.08E−05 9.08E−07 TGI 2.18E−05 2.18E−05 2.02E−05 1.34E−05 1.15E−05 2.18E−05 2.18E−05 4.02E−06 LC50 2.18E−05 2.18E−05 2.18E−05 2.18E−05 2.18E−05 2.18E−05 2.18E−05 2.18E−05 190 GI50 5.75E−08 4.86E−08 1.34E−07 1.02E−07 9.22E−08 9.15E−06 1.67E−07 7.18E−08 TGI 2.63E−07 2.97E−07 8.95E−07 1.20E−06 3.83E−07 1.07E−05 1.66E−06 7.12E−06 LC50 1.07E−05 1.97E−06 8.32E−06 6.42E−06 2.63E−06 1.07E−05 1.07E−05 1.07E−05 191 GI50 6.21E−08 2.44E−07 4.58E−07 5.88E−07 5.55E−07 3.63E−07 5.73E−07 1.29E−07 TGI 2.28E−07 1.48E−06 2.77E−06 6.96E−06 3.66E−06 1.17E−05 1.17E−05 1.69E−06 LC50 1.17E−05 5.36E−06 1.17E−05 1.17E−05 1.17E−05 1.17E−05 1.17E−05 9.49E−06 193 GI50 3.33E−08 1.18E−07 7.41E−08 1.36E−07 2.27E−07 1.01E−07 6.07E−07 1.23E−07 TGI 1.28E−07 4.91E−07 4.10E−07 4.32E−07 5.67E−07 4.97E−07 1.02E−05 3.99E−07 LC50 4.38E−07 5.49E−06 3.48E−06 9.59E−06 1.02E−05 1.02E−05 1.02E−05 3.91E−06 195 GI50 1.37E−08 2.21E−08 2.50E−08 4.17E−08 4.55E−08 4.41E−08 1.13E−07 1.17E−08 TGI 2.84E−08 8.64E−08 1.45E−07 2.15E−07 8.98E−08 1.59E−07 1.15E−06 4.09E−08 LC50 5.85E−08 3.65E−07 8.55E−07 2.74E−06 8.40E−07 1.94E−06 1.10E−05 1.70E−07 196 GI50 4.14E−07 3.98E−07 6.91E−07 8.49E−07 6.54E−07 6.75E−07 1.68E−06 2.34E−07 TGI 1.65E−06 1.45E−06 2.49E−06 3.85E−06 1.65E−06 8.99E−06 2.64E−05 6.14E−07 LC50 1.12E−05 9.78E−06 1.65E−05 2.64E−05 2.64E−05 2.64E−05 2.64E−05 1.50E−06 199 GI50 3.73E−09 1.58E−08 1.89E−08 3.13E−08 4.66E−08 3.26E−08 6.06E−08 1.12E−08 TGI 9.93E−09 8.92E−08 9.62E−08 1.59E−07 9.74E−08 1.51E−07 1.33E−06 5.55E−08 LC50 3.72E−08 3.92E−07 8.73E−07 1.22E−05 8.28E−06 7.04E−06 1.22E−05 2.81E−07 204 GI50 5.63E−09 7.90E−08 2.38E−08 1.80E−08 1.51E−07 2.68E−08 3.11E−08 7.48E−09 TGI 1.74E−08 2.29E−07 1.13E−07 1.09E−07 3.90E−07 2.43E−07 1.00E−06 2.03E−07 LC50 4.47E−08 5.97E−07 5.26E−07 9.52E−06 1.09E−06 7.14E−06 9.52E−06 5.96E−07 PANC-1 HT-29 LOVO LOVO-DOX HELA HELA-APL 135 GI50 2.11E−06 1.15E−06 2.69E−06 3.80E−06 TGI 5.53E−06 4.82E−06 8.76E−06 8.76E−06 LC50 8.76E−06 8.76E−06 8.76E−06 8.76E−06 169 GI50 1.52E−07 2.97E−06 1.59E−07 2.58E−07 5.57E−08 7.05E−08 TGI 1.05E−05 1.05E−05 4.18E−07 9.22E−07 1.39E−06 5.30E−07 LC50 1.05E−05 1.05E−05 1.42E−06 105E−05 4.42E−06 2.97E−06 173 GI50 4.61E−07 6.04E−06 1.15E−06 5.02E−07 1.21E−07 4.44E−07 TGI 6.04E−06 6.04E−06 2.57E−06 6.04E−06 6.04E−06 6.04E−06 LC50 6.04E−06 6.04E−06 5.72E−06 6.04E−06 6.04E−06 6.04E−06 180 GI50 1.30E−07 1.11E−05 5.35E−07 1.04E−07 8.06E−08 1.30E−07 TGI 1.11E−08 1.11E−05 2.01E−06 4.98E−07 1.99E−06 1.21E−06 LC50 1.11E−05 1.11E−05 8.51E−06 1.11E−05 1.11E−05 1.11E−05 181 GI50 8.64E−08 3.06E−06 3.31E−07 9.45E−08 4.78E−08 6.67E−08 TGI 2.09E−06 1.05E−05 1.10E−06 4.70E−07 6.32E−07 4.77E−07 LC50 1.05E−05 1.05E−05 1.05E−05 1.05E−05 1.05E−05 8.14E−06 182 GI50 4.85E−06 2.13E−05 2.13E−05 1.21E−06 8.52E−06 4.46E−06 TGI 2.13E−05 2.13E−05 2.13E−05 2.13E−05 2.13E−05 2.13E−05 LC50 2.13E−05 2.13E−05 2.13E−05 2.13E−05 2.13E−05 2.13E−05 188 GI50 5.56E−06 2.18E−05 4.50E−06 1.30E−06 1.05E−06 9.84E−07 TGI 2.18E−05 2.18E−05 2.18E−05 2.18E−05 3.69E−06 3.16E−05 LC50 2.18E−05 2.18E−05 2.18E−05 2.18E−05 2.18E−05 1.58E−05 190 GI50 3.76E−07 2.99E−06 2.85E−07 3.10E−07 8.33E−08 8.99E−08 TGI 3.50E−06 1.07E−05 1.20E−06 1.37E−06 1.31E−06 1.72E−06 LC50 1.07E−05 1.07E−05 5.46E−06 7.68E−06 3.93E−06 5.20E−06 191 GI50 1.27E−06 6.18E−06 5.13E−07 7.85E−07 9.48E−07 9.40E−07 TGI 9.66E−06 1.17E−05 1.70E−06 1.17E−05 2.96E−06 2.52E−06 LC50 1.17E−05 1.17E−05 7.90E−06 1.17E−05 8.63E−06 5.96E−06 193 GI50 2.78E−07 1.02E−05 3.50E−07 3.13E−07 3.50E−07 7.97E−07 TGI 3.60E−06 1.02E−05 8.47E−07 1.02E−05 1.02E−05 1.02E−05 LC50 1.02E−05 1.02E−05 1.02E−05 1.02E−05 1.02E−05 1.02E−05 195 GI50 5.69E−08 2.15E−06 1.50E−07 1.06E−07 5.13E−08 8.76E−08 TGI 3.63E−07 4.31E−06 3.42E−07 5.00E−07 2.04E−07 2.34E−07 LC50 3.11E−06 8.62E−06 7.83E−07 2.50E−06 6.33E−07 5.36E−07 196 GI50 9.57E−07 8.46E−06 7.99E−07 5.17E−07 3.51E−07 1.17E−07 TGI 7.25E−06 2.64E−05 3.53E−06 2.64E−05 1.93E−06 5.11E−07 LC50 2.64E−05 2.64E−05 2.64E−05 2.64E−05 2.64E−05 1.92E−06 199 GI50 3.69E−08 2.14E−06 1.28E−07 7.37E−08 2.99E−08 7.18E−08 TGI 2.76E−07 6.71E−06 3.35E−07 4.99E−07 1.99E−07 2.42E−07 LC50 8.08E−06 1.22E−05 8.79E−07 1.22E−05 6.51E−07 5.74E−07 204 GI50 4.47E−08 1.40E−06 1.13E−07 1.60E−07 3.69E−08 8.88E−08 TGI 1.71E−07 3.80E−06 2.28E−07 6.34E−06 1.72E−07 2.35E−07 LC50 3.92E−07 9.52E−06 4.63E−07 9.52E−06 4.52E−07 5.94E−07 DU-145 LN-caP IGROV IGROV-ET SK-BR-3 MEL-28 A-549 K-562 206 GI50 2.18E−08 7.52E−08 5.27E−08 2.97E−08 7.01E−08 4.40E−08 5.58E−08 3.11E−08 TGI 3.99E−08 4.75E−07 1.94E−07 1.90E−07 4.51E−07 1.69E−07 4.10E−07 1.11E−06 LC50 7.25E−08 3.33E−06 5.89E−07 5.89E−07 2.89E−06 1.04E−06 2.77E−06 4.23E−06 207 GI50 2.30E−08 1.27E−07 7.63E−08 4.36E−08 2.02E−07 4.44E−08 9.14E−08 9.20E−08 TGI 4.16E−08 9.44E−07 3.18E−07 2.86E−07 1.07E−06 2.45E−07 1.02E−06 3.06E−06 LC50 7.48E−08 4.10E−06 1.55E−06 2.38E−06 4.21E−06 4.60E−06 3.55E−06 9.26E−06 208 GI50 8.78E−06 1.46E−06 8.78E−06 8.78E−06 5.01E−06 8.78E−06 8.78E−06 TGI 8.78E−06 2.91E−06 8.78E−06 8.78E−06 7.97E−06 8.78E−06 8.78E−06 LC50 8.78E−06 5.83E−06 8.78E−06 8.78E−06 8.78E−06 8.78E−06 8.78E−06 209 GI50 4.29E−09 1.58E−08 3.37E−08 3.17E−08 4.78E−08 3.28E−08 4.28E−08 TGI 1.19E−08 5.60E−08 1.24E−07 7.01E−08 6.95E−08 8.39E−08 1.95E−07 LC50 4.04E−08 2.33E−07 4.77E−07 3.45E−07 1.01E−07 4.80E−07 7.13E−07 210 GI50 1.81E−07 3.17E−07 1.45E−07 3.97E−07 4.77E−07 6.12E−07 TGI 3.39E−07 1.06E−06 3.66E−07 1.36E−06 3.94E−06 2.13E−06 LC50 6.31E−07 3.65E−06 9.20E−07 4.59E−06 1.04E−05 6.17E−06 221 GI50 4.31E−08 7.22E−08 8.19E−08 6.48E−08 1.53E−06 1.06E−07 8.02E−08 8.83E−08 TGI 2.57E−07 5.02E−06 2.01E−05 2.01E−05 1.29E−05 2.01E−05 2.01E−05 2.01E−05 LC50 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 2.01E−05 233 GI50 7.83E−08 1.26E−07 4.99E−08 1.07E−07 1.69E−07 3.12E−07 9.58E−08 TGI 1.80E−07 5.57E−07 1.42E−07 7.21E−07 4.75E−06 2.06E−06 1.09E−06 LC50 1.95E−05 3.72E−06 1.95E−05 1.95E−05 1.95E−05 1.95E−05 8.61E−06 238 GI50 4.58E−08 2.13E−06 7.99E−08 4.13E−08 1.53E−07 5.26E−08 4.58E−08 4.33E−08 TGI 9.65E−08 1.65E−05 2.82E−07 1.44E−07 1.61E−06 3.51E−07 4.38E−07 2.47E−07 LC50 1.65E−05 1.65E−05 1.26E−06 1.65E−05 1.65E−05 1.25E−05 1.65E−05 1.20E−06 239 GI50 5.83E−08 2.42E−07 1.61E−07 2.06E−07 2.03E−07 1.64E−07 7.86E−08 TGI 1.31E−07 3.38E−06 4.05E−06 1.99E−06 2.60E−06 1.55E−05 2.16E−07 LC50 1.32E−06 1.65E−05 1.65E−05 1.65E−05 1.65E−05 1.65E−05 5.29E−07 240 GI50 6.06E−08 4.09E−06 1.36E−07 8.24E−08 5.93E−08 9.97E−08 1.26E−07 1.24E−07 TGI 1.34E−07 1.65E−05 5.22E−07 3.66E−07 3.43E−07 3.81E−07 4.70E−07 4.50E−07 LC50 1.65E−05 1.65E−05 3.64E−06 1.29E−05 1.65E−05 1.44E−06 1.51E−06 2.09E−06 PANC-1 HT-29 LOVO LOVO-DOX HELA HELA-APL 206 GI50 1.37E−07 1.32E−06 1.67E−07 2.94E−07 5.05E−08 8.34E−08 TGI 1.27E−06 1.05E−05 3.31E−07 1.01E−06 3.30E−07 5.23E−07 LC50 6.25E−06 1.05E−05 6.60E−07 6.37E−06 1.93E−06 2.82E−06 207 GI50 1.27E−07 3.77E−06 1.89E−07 3.16E−07 4.22E−08 7.79E−08 TGI 2.37E−06 9.26E−06 4.05E−07 2.85E−06 1.23E−06 1.50E−06 LC50 9.26E−06 9.26E−06 8.68E−07 9.26E−06 3.34E−06 3.71E−06 208 GI50 5.92E−06 8.78E−06 8.78E−06 8.78E−06 2.08E−07 1.54E−07 TGI 8.78E−06 8.78E−06 8.78E−06 8.78E−06 6.18E−07 3.85E−07 LC50 8.78E−06 8.78E−06 8.78E−06 8.78E−06 2.31E−06 1.14E−06 209 GI50 6.74E−08 3.38E−06 1.19E−07 1.37E−07 6.62E−08 8.27E−08 TGI 2.82E−07 9.99E−06 2.51E−07 4.88E−07 2.69E−07 2.21E−07 LC50 1.03E−06 1.05E−05 5.27E−07 2.60E−06 1.01E−06 5.17E−07 210 GI50 3.98E−07 7.09E−06 2.05E−06 2.43E−06 6.11E−07 5.16E−07 TGI 1.51E−06 1.04E−05 3.99E−06 6.04E−06 5.54E−06 3.08E−06 LC50 4.87E−06 1.04E−05 7.76E−06 1.04E−05 1.04E−05 1.04E−05 221 GI50 7.86E−08 2.01E−05 2.41E−06 2.83E−06 5.64E−08 7.96E−08 TGI 2.01E−05 2.01E−05 6.20E−06 1.42E−05 2.01E−05 2.01E−05 LC50 2.01E−05 2.01E−05 1.59E−05 2.01E−05 2.01E−05 2.01E−05 233 GI50 3.41E−07 1.95E−05 5.45E−07 1.05E−06 1.91E−07 4.17E−07 TGI 1.95E−05 1.95E−05 1.31E−06 1.54E−05 3.82E−06 3.15E−06 LC50 1.95E−05 1.95E−05 1.95E−05 1.95E−05 1.60E−05 1.43E−05 238 GI50 2.54E−08 2.65E−06 5.70E−08 2.26E−06 5.22E−08 1.57E−07 TGI 2.50E−07 1.06E−05 2.26E−07 8.98E−07 1.31E−07 5.16E−07 LC50 1.61E−06 1.65E−05 1.29E−06 1.65E−05 1.24E−06 1.76E−06 239 GI50 2.32E−07 1.08E−05 6.92E−07 8.71E−07 2.34E−07 6.56E−07 TGI 1.10E−05 1.65E−05 2.87E−06 1.65E−05 3.61E−06 4.18E−06 LC50 1.65E−05 1.65E−05 1.65E−05 1.65E−05 1.65E−05 1.65E−05 240 GI50 7.05E−08 1.64E−06 1.70E−07 4.61E−07 5.75E−08 3.41E−07 TGI 6.82E−07 1.59E−05 5.09E−07 1.15E−06 1.49E−07 1.40E−06 LC50 8.86E−06 1.65E−05 1.52E−06 1.65E−05 1.40E−06 7.61E−06

Example 3 Topoisomerase I Inhibition

The marine alkaloid lamellarin D (LAM-D, 3) has been recently characterized as a potent poison of human topoisomerase I endowed with remarkable cytotoxic activities against tumor cells. We report here the structure-activity relationship study in the LAM-D series.

Two groups of triester compounds incorporating various substituents on the three phenolic OH at positions 8, 14 and 20 of 6H-[1]benzopyrano[4′,3′:4,5]pyrrolo[2,1-a]isoquinolin-6-one pentacyclic planar chromophore typical of the parent alkaloid were tested as topoisomerase I inhibitors.

Compounds incorporating amino acid residues strongly promoted DNA cleavage by human topoisomerase I. LAM-D derivatives tri-substituted with leucine, valine, proline, phenylalanine or alanine residues, or a related amino side chain, stabilize topoisomerase I-DNA complexes. The DNA cleavage sites detected a T^(↓)G or C^(↓)G dinucleotides with these molecules were identical to that of LAM-D (3) but slightly different from those seen with camptothecin which stimulates topoisomerase I-mediated cleavage at T^(↓)G only.

With prostate (DU-145 and LN-CaP), ovarian (IGROV and IGROV-ET resistant to ecteinascidin-743) and colon (LoVo and LoVo-Dox cells resistant to doxorubicin) cancer cells (but not with HT29 colon carcinoma cells), the most cytotoxic compounds correspond to the most potent topoisomerase I poisons. The observed correlation between cytotoxicity and topoisomerase I inhibition strongly suggests that topoisomerase I-mediated DNA cleavage assays can be used as a guide to the development of superior analogs in this series.

Two assays, based on DNA relaxation and DNA cleavage (Bailly, C. DNA relaxation and cleavage assays to study topoisomerase I inhibitors. Methods Enzymol. 2001, 340, 610-623) were used evaluate the effects of the lamellarin analogs on the catalytic activity of human topoisomerase I.

In the first assay, a supercoiled plasmid DNA was relaxed with topoisomerase I in the absence or presence of the test compounds, each tested at 1 μM. DNA relaxation products were then resolved by gel electrophoresis on agarose gels containing ethidium bromide to stain the DNA. The alkaloid camptothecin, used as a positive control, strongly promotes DNA cleavage by topoisomerase I. Similarly, the intensity of the band corresponding to nicked DNA is significantly amplified in the presence of LAM-D (3) indicating that this natural product also stabilizes DNA-topoisomerase I covalent complexes. This functional assay is useful to identify the topoisomerase I poisons among the various analogs synthesized. The analogous compounds with a 5-6 saturated bond (11, 22, 108, 109, 139) were inactive in this assay.

Different cationic groups, mostly amino acid residues, were incorporated at the three phenoxy positions of LAM-D. A marked inhibition of topoisomerase I was observed with the positively charged molecules 40 (Ala), 39 (Leu), 36 (Val), 33 (Pro) and 25 (Phe) but not with the corresponding NH-Boc derivatives or the non-planar C5-C6 analogues. The Phe derivative is significantly less potent than the other amino acid derivatives which are all more or less equally effective at inhibiting topoisomerase I. The stereospecificity was investigated with the Val derivatives for which we compared the activity of the (L) (36, 38, 135, 144) and (D) (17, 32, 34, 122) isomers but there was no difference between the two series. Compound 17 and 36 both stimulated DNA cleavage by the enzyme. No effect was observed with the Boc-protected analogs in the C5-C6 double stranded (38, 122) or C5-C6 single-stranded (32, 34, 135, 144) series. The amino compounds 24 and 169 were also found to inhibit topoisomerase I.

Concentration-dependent measurements were performed with each of the positive compounds identified and a few representative gels comparing the anti-topoisomerase I activity of LAM-D (3) with the three analogues Val(D) (17), Pro (33) and the amino compound 169 were done. This later compound is equally efficient to (3) in terms of stimulation of DNA cleavage by topoisomerase I. In all cases, the dose-response analysis confirmed that the cationic LAM-D analogues potently inhibit the enzyme.

It is clear that the introduction of an amino acid functionality on the phenolic OH groups at positions 8, 14 and 20 of LAM-D (3) is not detrimental to topoisomerase I inhibition. The extent of topoisomerase I-mediated DNA cleavage is fully maintained when a Leu, Val, Ala or Pro residue is incorporated on the LAM-D skeleton whereas a non charged group abolishes the anti-topoisomerase I activity. A phenylalanine residue is much less favorable than a proline or an alanine residue for example. The observations that the incorporation of a cationic group promoted topoisomerase I inhibition suggested that the enhanced capacity of the drugs to bind to DNA could be responsible for a better enzyme inhibition.

A second assay, based on the cleavage of a radiolabeled DNA substrate by topoisomerase I, was used to confirm that the cationic lamellarin derivatives do effectively function as topoisomerase I poisons. A 117-bp DNA restriction fragment uniquely end-labeled at the 3′ end was subjected to cleavage by topoisomerase I in the presence of the different compounds and the resulting DNA cleavage products were resolved on sequencing-type polyacrylamide gels. The advantage of this assay is to detect the cleavage sites and to locate their positions with nucleotide resolution, providing thus information on the site selectivity of cleavage.

The reference drug CPT products three sites at nucleotide positions 26, 48 and 81 which all three correspond to T^(↓)G sites. Cleavage at TG sites in the presence of CPT is believed to result from the interaction of topoisomerase I with the T residue combined with the stacking of the CPT molecule with the adjacent G residue. A fourth weak site can be detected at the top of the gels (T^(↓)G107).

The sequence selectivity profiles are slightly different with the lamellarin analogs. LAM-D is less efficient than CPT for topoisomerase I-mediated DNA cleavage at sites T^(↓)G48 and T^(↓)G81 but it induces an additional cleavage site at C^(↓)G73. This likely reflects a different mode of interaction with the topoisomerase I-DNA covalent complexes.

Cleavage profiles identical to that of 3 were obtained with the cationic derivatives such as 39 (Leu), 36 (Val), 33 (Pro) and 25 (Phe) but not with the corresponding NH-Boc derivatives or the non-planar C5-C6 analogue. The amino compounds 169 and 7 were also found to stimulate DNA cleavage by the enzyme and here also we found no difference between the (L) (36) and (D) (17) Val isomers. The results are thus entirely consistent with those obtained by the relaxation assay and therefore validate the conclusion that the cationic lamellarin derivatives potently inhibit topoisomerase I. 

1. A compound of the general formula III:

wherein X is selected from the group consisting of N, O and S; wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₉ and R₉ are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO₂R′, NHR′, N(R′)₂, N═R′, NHCOR′, N(COR′)₂, NHSO₂R′, NO₂, PO(R′)₂, PO₂R′, C(═O)H, C(═O)R′, CO₂H, CO₂R′, OPO(R′)₂, OPO₂R′, OC(═O)H, OC(═O)R′, C(═O)R′, N═C(R′)₂, substituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₁-C₁₂ haloalkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, substituted or unsubstituted C₂-C₁₂ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and substituted or unsubstituted heteroaromatic; wherein each of the R′ groups is independently selected from the group consisting of H, OH, NO₂, NH₂, SH, CN, halogen, ═O, C(═O)H, C(═O)CH₃, CO₂H, C(═O)R′, substituted or unsubstituted C₁-C₁₈ alkyl, substituted or unsubstituted C₂-C₁₈ alkenyl, substituted or unsubstituted C₂-C₁₈ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C₁-C₁₈ alkoxyl, substituted or unsubstituted C₁-C₁₈ aminoalkyl, substituted or unsubstituted C₁-C₁₈ aminoacid, substituted or unsubstituted C₁-C₁₈ thioalkyl, substituted or unsubstituted C₁-C₁₈ alkylsulfinyl, substituted or unsubstituted C₁-C₁₈ alkylsulfonyl; wherein the pairs of groups R₁ and R₂, R₂ and R₃, R₃ and R₄, R₃ and R₉, R₄ and R₉, R₉ and R₅, R₉ and R₆, or R₆ and R₇, R₇ and R₈ may be joined into a carbocyclic or heterocyclic ring system; and the dotted line represents an single or double bond; or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof; with the proviso that the compounds are not known lamellarins.
 2. A compound according to claim 1 characterized in that it has formula IV:

wherein R₁-R₈ are as defined above and R′₂-R′₆ have the same definitions as for R₁-R₈ above; or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof.
 3. A compound according to claim 1 or 2 characterized in that X is preferably O or N.
 4. A compound according to claim 1 or 2 characterized in that X is O.
 5. A compound according to any of claims 1 to 4 characterized in that the dotted line is a double bond.
 6. A compound according to any of claims 1 to 5 characterized in that each of R₁-R₈ is independently selected from H, OR′, OC(═O)R′.
 7. A compound according to any of claims 1 to 6 characterized in that R₃ is selected from the group consisting of H, OH, alkoxy, preferably methoxy.
 8. A compound according to any of claims 1 to 6 characterized in that R₄, R₅, R₆ and R₈ are each independently selected from the group consisting of H or alkoxy.
 9. A compound according to claim 8 characterized in that R₄, R₅ and R₈ are H.
 10. A compound according to any of claims 1 to 5 characterized in that R₁, R₂ and R₇ are each independently selected from the group consisting of H, OH, alkoxy, OC(═O)R′, OSO₂R′, OPO(R′)₂, O-alkyl, NO₂, NH₂.
 11. A compound according to claim 10 characterized in that R₁, R₂ and R₇ are OC(═O)R′ wherein R′ is a substituted or unsubstituted aminoacid or aminoacids chain, preferably with a cationic group.
 12. A compound according to any of claims 2 to 11 characterized in that R′₂, R′₃ and R′₆ are each independently selected from the group consisting of H or alkoxy, preferably H.
 13. A compound according to any of claims 2 to 12 characterized in that R′₅ is selected from the group consisting of H or alkoxy, preferably methoxy.
 14. A compound according to any of claims 2 to 13 characterized in that R′₄ is selected from the group consisting of H, OH, alkoxy, OC(═O)R′, SO₂R′, PO(R′)₂, Alkyl, NO₂, NH₂.
 15. A compound according to claim 14 characterized in that R′₄ is C(═O)R′ wherein R′ is a substituted or unsubstituted aminoacid or aminoacids chain, preferably with a cationic group.
 16. A compound according to any of the preceding claims characterized in that at least one of R₁-R₈ and R′₂-R′₆ is not H, OH, OCH₃, SO₃Na, preferably at least two are not H, OH, OCH₃, SO₃Na.
 17. A pharmaceutical composition comprising a compound as defined in any of claims 1-16 or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof, and a pharmaceutically acceptable diluent or carrier.
 18. The use of a compound as defined in any of claims 1 to 16 or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof in the preparation of a medicament.
 19. A method of treating a tumour which comprises administering an effective amount of a compound as defined in any of claims 1 to 16 or a pharmaceutically acceptable salt, derivative, prodrug or stereoisomer thereof.
 20. The use of compounds as defined in any of claims 1 to 16 or pharmaceutically acceptable salts, derivatives, prodrugs or stereoisomers thereof as topoisomerase I inhibitors. 